NSD2 Antibody

What is NSD2 and why is it important in cancer research?

NSD2 (Nuclear Receptor Binding SET Domain Protein 2), also known as MMSET or WHSC1, is a SET domain-containing histone methyltransferase that catalyzes the methylation of histone H3 at lysine 36. It has gained significant attention in cancer research due to its overexpression in multiple cancer types. NSD2 is implicated in several cancers, including triple-negative breast cancer (TNBC), prostate cancer, neuroblastoma, carcinomas of the stomach and colon, small-cell lung cancers, and bladder cancers. Its overexpression is associated with tumor aggressiveness and poor patient survival, particularly in TNBC where high NSD2 expression correlates significantly with earlier disease-related death . At the molecular level, NSD2 regulates cancer cell survival, proliferation, and invasion by controlling important signaling pathways, including EGFR-AKT/STAT3 signaling in TNBC cells .

What applications are NSD2 antibodies validated for?

NSD2 antibodies have been validated for multiple applications crucial for epigenetic and cancer research:

When performing these applications, researchers should verify antibody specificity using appropriate controls, such as NSD2 knockdown samples, which have been demonstrated to show significant reduction in signal intensity in various assay formats .

How do I select the appropriate NSD2 antibody for my research?

Selection of the appropriate NSD2 antibody depends on several factors:

• Target specificity: Verify that the antibody specifically recognizes NSD2 (MMSET/WHSC1) and not other NSD family members.

• Application compatibility: Ensure the antibody is validated for your intended application (ChIP, WB, IF).

• Epitope recognition: Different antibodies recognize different epitopes of NSD2. For example, some antibodies are raised against the N-terminal region (amino acids 1-647) , while others target the C-terminal region (amino acids 959-1365) .

• Clonality: Monoclonal antibodies (like clone 29D1) offer high specificity for a single epitope, whereas polyclonal antibodies may provide stronger signals by recognizing multiple epitopes.

• Validation evidence: Review literature using the specific antibody and assess manufacturer validation data, including knockdown controls that demonstrate specificity .

For comprehensive studies, it may be beneficial to use multiple antibodies targeting different epitopes to confirm results and gain insights into potential isoform-specific functions.

What are the key considerations for optimizing ChIP experiments with NSD2 antibodies?

Chromatin immunoprecipitation (ChIP) with NSD2 antibodies requires careful optimization:

• Chromatin preparation: Use formaldehyde crosslinking (1% for 10 minutes at room temperature) followed by sonication to produce DNA fragments of 200-500 bp.

• Antibody amount: Most protocols recommend 2-5 μg of NSD2 antibody per ChIP reaction with chromatin from approximately 1×10^7 cells .

• Controls: Include:

  • Input control (untreated chromatin)

  • IgG control (non-specific antibody)

  • Positive control (antibody against a well-established histone mark)

  • Biological validation using NSD2 knockdown or knockout cells

• Target validation: Design primers for qPCR that target gene body regions known to be associated with NSD2 binding.

• Cross-reactivity assessment: Evaluate potential cross-reactivity with other SET domain proteins by comparing ChIP-seq profiles with published datasets.

For example, validated ChIP protocols have successfully used 4 μg of NSD2 antibody per reaction with chromatin from 1×10^7 KMS11 cells, with specificity confirmed using NSD2 knockout cells as negative controls .

How can I troubleshoot weak or non-specific signals in Western blots using NSD2 antibodies?

Troubleshooting weak or non-specific signals when using NSD2 antibodies in Western blotting:

• Protein extraction optimization:

  • Use RIPA buffer supplemented with protease inhibitors and phosphatase inhibitors

  • Include 1-2 mM PMSF to prevent proteolytic degradation

  • Consider adding deacetylase inhibitors (5-10 mM sodium butyrate)

• Loading and transfer considerations:

  • Load adequate protein (40-80 μg of whole cell lysate)

  • Run the gel long enough to resolve high molecular weight proteins (NSD2 is approximately 180 kDa)

  • Use wet transfer for large proteins (overnight at 30V at 4°C)

• Antibody incubation optimization:

  • Test different antibody concentrations (0.2-1 μg/ml)

  • Extend primary antibody incubation to overnight at 4°C

  • Reduce background by using 5% BSA instead of milk for blocking

• Validation strategies:

  • Include positive controls (cell lines known to express NSD2, like KMS11)

  • Include negative controls (NSD2 knockdown samples)

  • Test multiple NSD2 antibodies targeting different epitopes

If bands appear at unexpected molecular weights, consider the presence of post-translational modifications, degradation products, or alternative splicing variants of NSD2.

What methodological approaches are effective for studying NSD2 methyltransferase activity in vitro?

To study NSD2 methyltransferase activity in vitro, researchers can employ several methodological approaches:

• Peptide SPOT array methylation assays: These have been used to investigate substrate sequence specificity of NSD2, revealing strong readout of residues between G33 (-3) and P38 (+2) on the H3K36 sequence .

• Radiolabeled methyl transfer assays:

  • Purified recombinant NSD2 (typically the catalytic SET domain)

  • Substrate peptides or recombinant histones

  • ^3H-labeled S-adenosyl methionine (SAM) as methyl donor

  • Quantification via scintillation counting

• MALDI-TOF mass spectrometry:

  • Allows direct detection of methylation state changes

  • Can determine mono-, di-, or trimethylation states

  • Provides precise molecular mass shifts

• Antibody-based detection methods:

  • Western blotting with antibodies specific to methylated products

  • ELISA-based assays for quantitative assessment

• Super-substrate approach: Researchers have developed an engineered super-substrate (ssK36) that is methylated approximately 100-fold faster by NSD2 than the natural H3K36 substrate, providing a valuable tool for enhanced activity measurements .

When designing these experiments, it's crucial to include appropriate controls such as catalytically dead NSD2 mutants and to consider the influence of reaction conditions (pH, salt concentration, temperature) on enzymatic activity.

How is NSD2 antibody-based research contributing to our understanding of Triple-Negative Breast Cancer (TNBC)?

NSD2 antibody-based research has provided critical insights into the role of NSD2 in TNBC:

• Expression correlation with clinical outcomes: Immunohistochemistry using NSD2 antibodies has revealed that 62% of TNBC tumors show strong NSD2 expression compared to only 27% of non-TNBC tumors (p<0.0001). Kaplan-Meier analysis demonstrated that NSD2 protein overexpression in TNBC is significantly associated with earlier disease-related death .

• Molecular pathway identification: Through immunoblotting and immunoprecipitation studies, researchers have discovered that NSD2 regulates TNBC cell survival and invasion by directly controlling the expression and signaling of ADAM9 and EGFR. NSD2 silencing significantly inhibits EGFR and ADAM9 protein expression and reduces both total and phosphorylated forms of EGFR .

• Cellular function elucidation: Antibody-based detection following NSD2 silencing showed:

  • Decreased proliferation in MB-231 and MB-436 TNBC cell lines

  • Increased apoptosis as measured by TUNEL assay

  • Inhibited invasion capacity in Matrigel transwell assays

  • Reduced cell migration in wound-healing assays

• Signaling pathway impact: Western blotting with phospho-specific antibodies revealed that NSD2 silencing leads to marked reduction in AKT and STAT3 activation in TNBC cells, suggesting NSD2 is required for the hyperactivation of the EGFR-AKT/STAT3 signaling pathway .

These findings collectively suggest NSD2 as a potential therapeutic target for TNBC, with antibody-based methods providing crucial validation of its role and mechanism.

What are the methodological considerations when using NSD2 antibodies to study its non-histone substrates?

When investigating NSD2's non-histone substrates, researchers should consider several methodological approaches:

• Substrate identification strategies:

  • Immunoprecipitation with NSD2 antibodies followed by mass spectrometry

  • In vitro methylation assays using recombinant NSD2 and candidate substrate proteins

  • Peptide array screening based on substrate specificity profiles

• Validation of methylation:

  • Generation of methylation-specific antibodies for the identified sites

  • Mass spectrometry confirmation of methylation sites

  • Mutagenesis of predicted methylation sites (lysine to arginine substitutions)

• Cellular verification:

  • Co-immunoprecipitation experiments to confirm NSD2-substrate interactions

  • Methylation detection in cells with NSD2 overexpression or knockdown

  • Functional assays to determine the impact of substrate methylation

Recent research has identified ATP-dependent helicase (ATRX) K1033 and Fanconi anemia group M (FANCM) protein K819 as NSD2 protein substrates, demonstrating methylation both in vitro and in cells . These discoveries were made possible by combining specificity profiling of NSD2 with targeted validation assays.

To ensure specificity when studying non-histone substrates, researchers should:

• Use multiple NSD2 antibodies targeting different epitopes

• Include catalytically inactive NSD2 mutants as negative controls

• Compare results with other methyltransferases to confirm specificity to NSD2

How can NSD2 antibodies be used to investigate the role of NSD2 in NF-κB signaling pathways?

NSD2 antibodies are valuable tools for investigating the multilevel regulation of NF-κB signaling by NSD2:

• Co-immunoprecipitation studies: NSD2 has been immunoprecipitated from cell extracts of KPC1199 and PANC1 cells (treated with TNF-α) and immunoblotted with antibodies against NF-κB components to demonstrate direct interaction between NSD2 and the DNA binding domain of p65 .

• Chromatin binding analysis:

  • Electrophoretic mobility shift assays (EMSA) have shown that NSD2 depletion enhances NF-κB binding to the κB site, while NSD2 overexpression decreases it .

  • ChIP-seq assays have revealed that NSD2 overexpression results in reduced intensity of p65 binding signals around gene promoter regions .

  • ChIP-qPCR has been used to validate p65 binding at specific promoters of target genes .

• Functional impact assessment:

  • NSD2 antibodies can be used in Western blotting to measure changes in NF-κB pathway component expression and activation following NSD2 manipulation.

  • Immunofluorescence can detect changes in nuclear translocation of NF-κB components upon NSD2 overexpression or knockdown.

  • Proximity ligation assays (PLA) can visualize and quantify interactions between NSD2 and NF-κB components in situ.

When designing experiments to study NSD2-NF-κB interactions, researchers should consider both direct protein-protein interactions and the potential effect of NSD2-mediated histone methylation on chromatin accessibility at NF-κB target genes, employing appropriate controls such as NSD2 catalytic mutants to distinguish between these mechanisms.

What are common pitfalls when using NSD2 antibodies and how can they be addressed?

Common pitfalls when working with NSD2 antibodies include:

• Cross-reactivity with other NSD family members:

  • Problem: NSD1, NSD2, and NSD3 share significant homology in their SET domains.

  • Solution: Validate antibody specificity using NSD2 knockdown or knockout samples . Using antibodies targeting unique regions outside the SET domain can enhance specificity.

• Isoform detection issues:

  • Problem: NSD2 has multiple isoforms that may not all be recognized by a single antibody.

  • Solution: Review the immunogen sequence of the antibody to determine which isoforms it can detect. Consider using multiple antibodies targeting different regions if comprehensive isoform detection is needed.

• Fixation sensitivity in immunofluorescence:

  • Problem: Some epitopes may be masked by certain fixation methods.

  • Solution: Compare different fixation protocols (e.g., formaldehyde vs. methanol). Validated protocols show success with formaldehyde-fixed U2OS cells using 2 μg/ml of NSD2 antibody .

• High background in ChIP experiments:

  • Problem: Non-specific binding can obscure true signals.

  • Solution: Increase washing stringency, optimize antibody concentration (typically 2-5 μg per ChIP), and include appropriate controls including NSD2 knockout samples .

• Detecting post-translational modifications:

  • Problem: PTMs may affect antibody binding or create unexpected banding patterns.

  • Solution: Use phosphatase treatments on samples to determine if phosphorylation affects antibody recognition.

• Batch-to-batch variation:

  • Problem: Different lots of the same antibody may perform differently.

  • Solution: Record lot numbers used in successful experiments and validate new lots against previous ones before committing to large studies.

How can researchers validate the specificity of NSD2 antibodies in their experimental systems?

Validating the specificity of NSD2 antibodies is crucial for ensuring reliable experimental results. Recommended validation approaches include:

• Genetic knockdown or knockout controls:

  • siRNA knockdown of NSD2 should result in reduced signal intensity in Western blots, immunofluorescence, and ChIP experiments .

  • CRISPR/Cas9-mediated knockout cells provide the most stringent negative control .

• Blocking peptide competition:

  • Pre-incubate the antibody with excess immunizing peptide/protein

  • A specific antibody will show significantly reduced or eliminated signal

• Multiple antibody concordance:

  • Use different antibodies targeting distinct epitopes of NSD2

  • Consistent results across different antibodies increase confidence in specificity

• Recombinant protein controls:

  • Test antibody reactivity against purified recombinant NSD2

  • Include related proteins (NSD1, NSD3) to assess cross-reactivity

• Immunoprecipitation-Western blot validation:

  • Immunoprecipitate with one NSD2 antibody and blot with another

  • Confirms that both antibodies detect the same protein

• Mass spectrometry confirmation:

  • Immunoprecipitate NSD2 and verify its identity by mass spectrometry

  • Provides unambiguous identification of the detected protein

Published validation data shows the efficacy of these approaches. For example, Western blot validation of NSD2 antibody using KMS11 whole cell extract shows a clear band at 180 kDa that disappears in extracts from KMS11 cells with NSD2 knocked down via RNAi .

What are the key differences between antibodies targeting different domains of NSD2, and how do they impact experimental outcomes?

Different NSD2 antibodies target distinct domains of the protein, which can significantly impact experimental outcomes:

Impact on experimental outcomes:

• Isoform detection differences:

  • The NSD2 gene can produce multiple isoforms through alternative splicing

  • N-terminal antibodies typically detect most isoforms

  • Domain-specific antibodies may provide insights into isoform-specific functions

• Functional correlation variations:

  • SET domain-targeting antibodies may better correlate with methyltransferase activity

  • Antibodies against regulatory domains might better reflect activation state

• Epitope accessibility considerations:

  • In fixed samples or native protein conformations, certain epitopes may be masked

  • C-terminal epitopes may be more accessible in ChIP experiments due to protein orientation on chromatin

• Post-translational modification interference:

  • Antibodies targeting regions subject to PTMs may show reduced binding when the site is modified

  • This can be exploited to study regulation but may complicate interpretation of results

When designing experiments, researchers should select antibodies based on the specific research question. For total NSD2 detection, N-terminal antibodies are often preferred, while functional studies may benefit from SET domain-specific antibodies. For comprehensive characterization, using multiple antibodies targeting different domains provides the most complete picture.

How can NSD2 antibodies be leveraged in drug discovery research targeting NSD2 in cancer?

NSD2 antibodies are invaluable tools in drug discovery efforts targeting NSD2 in cancer:

• High-throughput screening (HTS) assay development:

  • NSD2 antibodies can be used in AlphaLISA or TR-FRET assays to detect methylation activity

  • These assays can screen compound libraries for inhibitors of NSD2 enzymatic activity

  • Western blotting with NSD2 antibodies can validate hits from primary screens by assessing effects on NSD2 expression and downstream targets

• Target engagement validation:

  • Cellular thermal shift assays (CETSA) using NSD2 antibodies can confirm direct binding of compounds to NSD2 in cells

  • Immunoprecipitation with NSD2 antibodies followed by drug competition assays can assess binding affinity

  • ChIP assays can determine if inhibitors disrupt NSD2 chromatin binding

• Pharmacodynamic (PD) biomarker development:

  • Antibodies against NSD2 and its substrate H3K36me2 can monitor target inhibition

  • Immunohistochemistry of patient samples can assess NSD2 expression levels to identify potential responders

  • Sequential tumor biopsies can be analyzed to confirm on-target activity during clinical trials

• Combination therapy rationale:

  • Pathway analysis following NSD2 inhibition can identify synergistic targets

  • For instance, research showing NSD2 regulation of EGFR-AKT/STAT3 signaling in TNBC suggests potential synergy with EGFR inhibitors

  • NSD2 antibodies can monitor pathway modulation in these combination approaches

• Resistance mechanism studies:

  • Antibody-based proteomics can identify adaptive responses to NSD2 inhibition

  • ChIP-seq with NSD2 antibodies can map changes in chromatin binding patterns in resistant cells

Given that NSD2 overexpression is associated with poor survival in TNBC and other cancers , developing effective inhibitors represents an important therapeutic opportunity, with antibody-based methods providing crucial tools throughout the drug discovery pipeline.

What are the considerations for using NSD2 antibodies in multiplex immunofluorescence or mass cytometry experiments?

When incorporating NSD2 antibodies into multiplex immunofluorescence or mass cytometry experiments, researchers should consider:

• Antibody compatibility and panel design:

  • Ensure primary antibodies are from different host species to avoid cross-reactivity

  • For mass cytometry, verify that metal conjugation doesn't interfere with NSD2 epitope recognition

  • When designing panels, include markers that provide biological context for NSD2 function (e.g., cell cycle markers, other epigenetic modifiers)

• Signal optimization strategies:

  • Titrate antibody concentrations to minimize background while maintaining specific signal

  • For IF, recommended starting dilutions of 1-2 μg/ml have been validated

  • Optimize fixation and permeabilization protocols to ensure epitope accessibility while preserving cellular morphology

• Controls for multiplex experiments:

  • Single-stain controls to assess spectral overlap and establish compensation

  • FMO (fluorescence minus one) controls to set accurate gating thresholds

  • Biological positive and negative controls (e.g., NSD2 knockdown cells)

  • Isotype controls to assess non-specific binding

• Subcellular localization considerations:

  • NSD2 primarily localizes to the nucleus, so nuclear segmentation is crucial

  • Co-staining with nuclear markers (DAPI, Hoechst) enables accurate quantification

  • Z-stack imaging may be necessary to capture the full nuclear volume

• Data analysis approaches:

  • Single-cell analysis allows correlation of NSD2 levels with other markers

  • Machine learning algorithms can identify cell subpopulations based on marker patterns

  • Spatial analysis can reveal relationships between NSD2-expressing cells and their microenvironment

These advanced approaches allow researchers to contextualize NSD2 expression and function within heterogeneous cell populations, providing insights into its role in complex biological systems like tumors.

How can researchers integrate ChIP-seq data generated with NSD2 antibodies with other epigenomic datasets?

Integrating NSD2 ChIP-seq data with other epigenomic datasets enables comprehensive understanding of NSD2's role in chromatin regulation:

• Multi-omics data integration strategies:

  • Compare NSD2 binding sites with histone modification patterns (H3K36me2, H3K4me3, H3K27ac)

  • Correlate with transcriptomic data (RNA-seq) to identify direct transcriptional targets

  • Overlap with chromatin accessibility data (ATAC-seq, DNase-seq) to assess impact on chromatin structure

  • Integrate with 3D genome organization data (Hi-C, ChIA-PET) to understand higher-order chromatin effects

• Computational analysis approaches:

  • Peak calling using MACS2 or similar algorithms to identify significant NSD2 binding sites

  • Motif analysis to identify DNA sequences enriched at NSD2 binding sites

  • Gene ontology and pathway analysis of NSD2-bound genes

  • Differential binding analysis between conditions (e.g., treatment vs. control)

• Validation experiments:

  • ChIP-qPCR to validate binding at specific loci of interest

  • Sequential ChIP (Re-ChIP) to determine co-occupancy with other factors

  • CUT&RUN or CUT&Tag as complementary approaches with potentially higher resolution

• Biological context considerations:

  • Cell type-specific binding patterns may reveal context-dependent functions

  • Treatment conditions (e.g., TNF-α stimulation) can reveal dynamic regulatory mechanisms

  • Disease state comparisons may identify pathological alterations in NSD2 function

• Visualization and interpretation tools:

  • Genome browsers (IGV, UCSC) for visualizing binding patterns

  • Heatmaps and aggregate plots to summarize binding relative to genomic features

  • Network analysis to identify cooperating factors and pathways

A powerful example from the literature shows how ChIP-seq analyses revealed that NSD2 overexpression results in reduction of p65 binding signal intensity around gene promoter regions, providing mechanistic insights into NSD2's role in suppressing NF-κB signaling . Such integrated approaches generate testable hypotheses about NSD2's functional roles in normal and disease states.

What emerging techniques might enhance the utility of NSD2 antibodies in epigenetic research?

Several emerging techniques promise to enhance the utility of NSD2 antibodies in epigenetic research:

• CUT&Tag and CUT&RUN technologies:

  • These techniques offer higher signal-to-noise ratios than traditional ChIP

  • Require fewer cells (as few as 1,000 compared to millions for ChIP-seq)

  • Can be performed in single cells to assess heterogeneity

  • NSD2 antibodies could be adapted to these protocols for improved chromatin profiling

• Proximity labeling approaches:

  • APEX2 or BioID fused to NSD2 can identify proximal interacting proteins

  • When combined with NSD2 antibodies for validation, provides comprehensive interactome mapping

  • Can reveal transient interactions missed by traditional immunoprecipitation

• Live-cell imaging of NSD2:

  • Nano-antibodies or intrabodies against NSD2 could enable live tracking

  • Reveals dynamic behavior and response to cellular stimuli

  • Can be combined with super-resolution microscopy for detailed localization studies

• Single-cell epigenomics:

  • Adaptation of NSD2 antibodies for single-cell CUT&Tag

  • Reveals cell-to-cell variation in NSD2 chromatin occupancy

  • Integration with single-cell transcriptomics for comprehensive understanding

• Targeted protein degradation tools:

  • NSD2 antibodies can validate degradation by PROTAC or molecular glue approaches

  • Enables acute depletion studies to distinguish direct from indirect effects

  • Provides alternative therapeutic strategy to enzymatic inhibition

These emerging techniques will allow researchers to gain increasingly sophisticated insights into NSD2 function, potentially revealing new therapeutic opportunities in cancer and other diseases where NSD2 dysregulation plays a role.

How might research on NSD2 non-histone substrates transform our understanding of its role in disease?

Research on NSD2 non-histone substrates could fundamentally transform our understanding of its role in disease:

• Expanded functional landscape:

  • Recent research has identified ATP-dependent helicase (ATRX) K1033 and Fanconi anemia group M (FANCM) protein K819 as NSD2 protein substrates in vitro and in cells

  • These discoveries suggest NSD2 function extends beyond histone modification to regulate diverse cellular processes

• Novel pathogenic mechanisms:

  • Methylation of non-histone proteins may alter their:

    • Stability and turnover rates

    • Protein-protein interaction capabilities

    • Subcellular localization

    • Enzymatic activities

  • These changes could represent previously unrecognized disease mechanisms

• Therapeutic implications:

  • Targeting NSD2's interaction with specific non-histone substrates might allow more precise interventions

  • Could enable selective disruption of pathogenic functions while preserving normal ones

  • May explain differential effectiveness of NSD2 inhibition across cancer types

• Methodological advances needed:

  • Development of substrate-specific methylation antibodies

  • Proteome-wide mapping of lysine methylation changes upon NSD2 modulation

  • Structural studies of NSD2 in complex with non-histone substrates

  • Functional characterization of methylation-deficient mutants of identified substrates

• Integration with cancer biology:

  • The discovery of NSD2's role in regulating ADAM9 and EGFR expression raises the possibility that direct methylation of components in these pathways might occur

  • NSD2's interference with NF-κB binding suggests potential direct methylation of NF-κB components

As research progresses, a more complete picture of NSD2's "methylome" may emerge, potentially revealing unexpected connections to disease processes and providing new opportunities for therapeutic intervention.

What unanswered questions about NSD2 should researchers prioritize investigating?

Several critical unanswered questions about NSD2 warrant prioritized investigation:

• Isoform-specific functions and localization:

  • How do different NSD2 isoforms contribute to normal development versus pathological states?

  • Do specific isoforms preferentially methylate certain substrates or localize to particular chromatin regions?

  • Can isoform-specific antibodies be developed to distinguish their unique roles?

• Regulatory mechanisms controlling NSD2 activity:

  • What post-translational modifications regulate NSD2 catalytic activity or substrate selection?

  • How is NSD2 recruitment to chromatin dynamically controlled in response to cellular signals?

  • What protein complexes contain NSD2, and how do they modulate its function?

• Comprehensive substrate profiling:

  • Beyond the recently identified ATRX K1033 and FANCM K819 , what is the complete repertoire of NSD2 non-histone substrates?

  • How does substrate specificity differ between NSD family members (NSD1, NSD2, NSD3)?

  • What is the functional significance of each methylation event?

• Therapeutic targeting strategies:

  • Can effective and selective NSD2 inhibitors be developed for cancer therapy?

  • Would targeting NSD2's catalytic activity or protein-protein interactions be more effective?

  • How can patient populations likely to respond to NSD2-targeted therapy be identified?

• Context-dependent roles in different cancer types:

  • Why is NSD2 particularly important in TNBC compared to other breast cancer subtypes ?

  • Does NSD2 drive similar or different pathways across cancer types where it is overexpressed?

  • Are there cancer-specific vulnerabilities created by NSD2 overexpression?

• Connection to broader epigenetic networks:

  • How does NSD2-mediated H3K36 methylation interact with other histone modifications?

  • What is the relationship between NSD2 activity and DNA methylation patterns?

  • How does NSD2 contribute to higher-order chromatin organization?

Addressing these questions will require innovative experimental approaches and integration of multiple technologies, with NSD2 antibodies remaining essential tools throughout this journey of discovery.