KDM6B Antibody, FITC conjugated

What is KDM6B and what biological functions does it regulate?

KDM6B/JMJD3 is a JmjC domain-containing histone demethylase that catalyzes the removal of silencing methyl groups on H3K27, thereby activating gene expression. By interacting with Set1/MLL methyltransferase, it also positively regulates the activating methylation of H3K4 . KDM6B regulates numerous biological processes including:

• Posterior development through HOX gene expression control

• Inflammatory responses and macrophage differentiation during inflammation

• Hematopoietic stem and progenitor cell function

• Neuronal maturation beyond early neuronal differentiation stages

• Development and function of intestinal intraepithelial lymphocytes (IELs), particularly TCRαβ+CD8αα+ IELs

What are the key specifications of commercially available KDM6B Antibody, FITC conjugated?

KDM6B Antibody, FITC conjugated is typically a rabbit polyclonal antibody targeting specific epitopes of the KDM6B/JMJD3 protein. Key specifications include:

• Conjugate: FITC (Fluorescein isothiocyanate) with excitation at 495 nm and emission at 519 nm

• Host: Rabbit

• Clonality: Polyclonal

• Isotype: IgG

• Reactivity: Validated for human and mouse samples, with predicted reactivity to rat (100%)

• Applications: Flow cytometry, immunocytochemistry/immunofluorescence, immunohistochemistry, western blot, ELISA, and dot blot

• Subcellular localization: Nucleus

How should KDM6B Antibody, FITC conjugated be stored and handled to maintain optimal activity?

For optimal performance and longevity of KDM6B Antibody, FITC conjugated:

• Store at -20°C or -80°C upon receipt

• Avoid repeated freeze-thaw cycles as this can compromise antibody function

• Protect from light due to the photosensitive nature of the FITC fluorophore

• When working with the antibody, maintain cold temperature (on ice) when possible

• Prepare aliquots for regular use to minimize freeze-thaw cycles

• For short-term storage (1-2 weeks), antibody can be kept at 4°C protected from light

• Follow manufacturer's specific recommendations for diluent composition (typically containing 50% glycerol, PBS pH 7.4, and preservatives like Proclin 300)

What are the optimal experimental conditions for using KDM6B Antibody, FITC conjugated in flow cytometry?

For flow cytometric applications using KDM6B Antibody, FITC conjugated:

• Cell preparation:

  • Harvest cells (~1×10⁶ cells per sample)

  • Fix with 4% paraformaldehyde (10 minutes at room temperature)

  • Permeabilize using 0.1% Triton X-100 or commercial permeabilization buffer (10 minutes)

• Antibody staining:

  • Block with 3-5% normal serum (from the same species as the secondary antibody)

  • Determine optimal antibody dilution experimentally (typically 1:50-1:200)

  • Incubate cells with diluted antibody for 30-60 minutes at room temperature in the dark

  • Wash 3 times with PBS containing 1% BSA or FBS

• Controls:

  • Include an unstained control

  • Include an isotype control (FITC-conjugated rabbit IgG)

  • Consider a blocking peptide control if available

• Instrument settings:

  • Use appropriate filter sets for FITC detection (excitation: 495 nm, emission: 519 nm)

  • Perform fluorescence compensation if multiple fluorophores are used

How can KDM6B Antibody, FITC conjugated be used in immunofluorescence studies of epigenetic regulation?

For immunofluorescence applications studying epigenetic regulation with KDM6B antibody:

• Sample preparation:

  • For adherent cells: culture on coverslips, fix with 4% paraformaldehyde (15 minutes), permeabilize with 0.25% Triton X-100 (10 minutes)

  • For tissue sections: deparaffinize, rehydrate, and perform antigen retrieval (citrate buffer pH 6.0)

• Immunostaining protocol:

  • Block with 5% normal serum in PBS containing 0.3% Triton X-100 (1 hour)

  • Dilute KDM6B Antibody, FITC conjugated appropriately (optimize experimentally)

  • Incubate overnight at 4°C in a humidified chamber

  • Wash 3 times with PBS (5 minutes each)

  • Counterstain nuclei with DAPI (1:1000 dilution, 5 minutes)

  • Mount with anti-fade mounting medium

• Co-staining options:

  • For epigenetic studies, consider co-staining with antibodies against:

    • Histone H3K27me3 (target of KDM6B demethylase activity)

    • RNA Polymerase II (marker of active transcription)

    • Other histone modifications (H3K4me3, H3K9me3)

• Advanced imaging:

  • Capture images using confocal microscopy for detailed subcellular localization

  • Perform z-stack imaging to fully visualize nuclear distribution

  • Use appropriate filters for FITC (excitation: 495 nm, emission: 519 nm)

  • Consider deconvolution for enhanced resolution of nuclear structures

What are the recommended protocols for using KDM6B Antibody, FITC conjugated in western blot applications?

For western blot applications using KDM6B Antibody, FITC conjugated:

• Sample preparation:

  • Extract nuclear proteins (as KDM6B is a nuclear protein)

  • Add protease and phosphatase inhibitors to lysis buffer

  • Determine protein concentration using Bradford or BCA assay

• Gel electrophoresis and transfer:

  • Load 20-40 μg protein per lane

  • Separate proteins using 8% SDS-PAGE (KDM6B is approximately 178 kDa)

  • Transfer to PVDF membrane (0.45 μm pore size recommended for large proteins)

• Immunoblotting:

  • Block membrane with 5% non-fat dry milk in TBS-T (1 hour at room temperature)

  • Dilute KDM6B Antibody, FITC conjugated appropriately (determine optimal dilution experimentally)

  • Incubate overnight at 4°C with gentle rocking

  • Wash 3 times with TBS-T (10 minutes each)

• Detection and visualization:

  • For FITC visualization: Use fluorescence imaging systems capable of detecting FITC

  • Expected molecular weight: ~178 kDa for full-length KDM6B protein

  • Include positive control samples (cell lines known to express KDM6B)

  • Consider loading controls (HDAC1, Lamin B1) for nuclear proteins

Note: While most antibodies for western blot are used with secondary antibodies, the FITC conjugation allows direct visualization with fluorescence imaging systems.

What are common troubleshooting strategies for weak or absent signal when using KDM6B Antibody, FITC conjugated?

When encountering weak or absent signals with KDM6B Antibody, FITC conjugated:

• Antibody-related issues:

  • Increase antibody concentration (titrate within recommended range)

  • Verify antibody integrity (avoid repeated freeze-thaw cycles)

  • Check storage conditions (improper storage can reduce activity)

  • Confirm the antibody hasn't expired or been compromised

• Sample preparation issues:

  • Ensure proper fixation and permeabilization for intracellular/nuclear staining

  • Optimize antigen retrieval conditions for tissue sections

  • Verify target protein expression in your sample type

  • For western blot, ensure complete transfer of high molecular weight proteins

• Technical considerations:

  • Optimize incubation time and temperature

  • Use fresh reagents, particularly fluorophore-sensitive applications

  • Protect from light to prevent photobleaching of FITC

  • Adjust exposure settings for imaging or detector sensitivity for flow cytometry

• Validation approaches:

  • Test antibody on positive control samples (e.g., cell lines known to express KDM6B)

  • Consider alternative detection methods (e.g., for western blot, try HRP-conjugated secondary antibody)

  • Verify KDM6B expression at mRNA level using RT-PCR

How can researchers distinguish between specific and non-specific staining when using KDM6B Antibody, FITC conjugated?

To distinguish between specific and non-specific staining:

• Essential controls:

  • Isotype control: Use FITC-conjugated rabbit IgG at the same concentration

  • Blocking peptide control: Pre-incubate antibody with immunizing peptide before staining

  • Negative control samples: Use cells/tissues known not to express KDM6B

  • Positive control samples: Use cells/tissues with validated KDM6B expression

• Pattern analysis:

  • Specific KDM6B staining should localize to the nucleus

  • Non-specific staining often appears as:

    • Uniform cytoplasmic signal

    • Cell membrane staining (KDM6B is not a membrane protein)

    • Signal in negative control samples

• Cross-validation techniques:

  • Compare with immunohistochemistry using non-FITC conjugated KDM6B antibody

  • Confirm knockdown efficiency with siRNA or shRNA against KDM6B

  • Perform dual staining with another KDM6B antibody recognizing a different epitope

• Technical approaches to reduce non-specific binding:

  • Increase blocking time/concentration

  • Pre-adsorb antibody with tissue homogenate

  • Optimize wash steps (increase number/duration)

  • Fine-tune antibody dilution

How can KDM6B Antibody, FITC conjugated be utilized to study the role of KDM6B in hematopoietic disorders?

For investigating KDM6B's role in hematopoietic disorders using FITC-conjugated antibody:

• Flow cytometric analysis of patient samples:

  • Isolate bone marrow hematopoietic stem and progenitor cells (HSPCs) using appropriate markers

  • Measure KDM6B expression levels in HSPCs from MDS and CMML patients compared to healthy controls

  • Create multiparameter panels to correlate KDM6B expression with disease markers and progenitor subpopulations

• Mechanistic studies on innate immune activation:

  • Analyze KDM6B expression in response to TLR ligand stimulation (e.g., LPS)

  • Compare expression of innate immune genes (e.g., S100a9) in cells with different KDM6B levels

  • Correlate KDM6B expression with H3K27me3 levels at promoters of innate immune genes

• Therapeutic intervention assessment:

  • Monitor KDM6B expression during treatment with GSK-J4 (KDM6B inhibitor)

  • Track changes in hematopoietic differentiation markers following KDM6B inhibition

  • Create experimental models that test the efficacy of KDM6B targeting in combination with standard MDS treatments

Research insight: Studies have shown that KDM6B is significantly overexpressed in bone marrow HSPCs of patients with MDS and CMML. Overexpression of KDM6B mediates activation of innate immune signals and plays a role in MDS and CMML pathogenesis. Pharmacologic inhibition of KDM6B with GSK-J4 has shown therapeutic potential in ameliorating ineffective hematopoiesis .

What methodologies can integrate KDM6B Antibody, FITC conjugated with ChIP-seq data to study epigenetic regulation?

Integrating KDM6B antibody immunofluorescence with ChIP-seq for comprehensive epigenetic studies:

• Combined IF-ChIP approach:

  • Use KDM6B Antibody, FITC conjugated for immunofluorescence to visualize nuclear localization

  • Perform ChIP-seq using KDM6B antibodies to identify genomic binding sites

  • Correlate KDM6B binding with H3K27me3 demethylation and gene activation

• Multi-omics integration workflow:

  • Step 1: Immunofluorescence with KDM6B Antibody, FITC conjugated to determine cell-type specific expression

  • Step 2: Cell sorting of positive populations

  • Step 3: ChIP-seq for KDM6B, H3K27me3, and H3K4me3 on sorted populations

  • Step 4: RNA-seq to correlate binding with gene expression changes

  • Step 5: Data integration using bioinformatics approaches

• Single-cell analysis techniques:

  • Combine flow cytometry using KDM6B Antibody, FITC conjugated with single-cell RNA-seq

  • Use computational approaches to integrate KDM6B protein levels with transcriptome data

  • Create pseudotime trajectories to understand KDM6B's role in cellular differentiation

• Validation experiments:

  • Use gene editing (CRISPR-Cas9) to modify KDM6B levels and assess changes in H3K27me3 landscape

  • Perform reporter assays at KDM6B binding sites identified by ChIP-seq

  • Conduct rescue experiments to confirm specificity of observed epigenetic changes

How can researchers study the dynamics of KDM6B interaction with other epigenetic regulators using the FITC-conjugated antibody?

To investigate KDM6B interactions with other epigenetic regulators:

• Advanced co-localization studies:

  • Perform multi-color immunofluorescence with KDM6B Antibody, FITC conjugated and antibodies against interacting partners

  • Use super-resolution microscopy (STED, STORM) for detailed nuclear co-localization analysis

  • Apply proximity ligation assay (PLA) to visualize and quantify protein interactions in situ

• Dynamic interaction analysis:

  • Implement live-cell imaging with KDM6B-FITC antibody in permeabilized cells

  • Perform fluorescence recovery after photobleaching (FRAP) to assess KDM6B mobility

  • Use fluorescence resonance energy transfer (FRET) between KDM6B-FITC and other labeled epigenetic factors

• Biochemical interaction studies complementing microscopy:

  • Conduct co-immunoprecipitation followed by western blot to validate interactions

  • Perform mass spectrometry analysis of KDM6B interactome under different conditions

  • Use chromatin immunoprecipitation (ChIP) with re-ChIP to identify genomic regions with co-bound factors

• Functional regulation studies:

  • Analyze the effect of KDM6B overexpression or inhibition on interacting partners

  • Study how stimuli that activate KDM6B (e.g., inflammatory signals) affect its interactions

  • Investigate post-translational modifications that regulate KDM6B interactions

Research insight: KDM6B has been shown to interact with the Set1/MLL methyltransferase complex to positively regulate H3K4 methylation in addition to its H3K27 demethylase activity, creating a coordinated epigenetic activation mechanism .

How can KDM6B Antibody, FITC conjugated be used to investigate intestinal immunology and colorectal cancer models?

For studying KDM6B in intestinal immunity and colorectal cancer:

• Analysis of intestinal intraepithelial lymphocytes (IELs):

  • Use flow cytometry with KDM6B Antibody, FITC conjugated to assess expression in different IEL subsets

  • Create a comprehensive panel including:

    • TCRαβ markers

    • CD8α and CD8β to differentiate CD8αα+ from CD8αβ+ IELs

    • KDM6B expression using the FITC-conjugated antibody

  • Compare expression in conventional vs. unconventional IELs

• Tumor microenvironment studies:

  • Perform immunofluorescence of intestinal tissue sections from cancer models

  • Co-stain with KDM6B Antibody, FITC conjugated and markers for:

    • T cell subsets (CD3, CD8)

    • Tumor cells (cytokeratins, β-catenin)

    • Proliferation markers (Ki67)

• Functional studies in mouse models:

  • Monitor KDM6B expression in APCMin/+ mice with or without IEL-specific Kdm6b deletion

  • Correlate KDM6B levels with intestinal tumorigenesis progression

  • Analyze expression of KDM6B-regulated genes (Bcl2, Gzmb, Fasl) in sorted IELs

Research insight: Kdm6b deficiency in IELs has been shown to aggravate tumorigenesis in the small intestine in APCMin/+ mice. Mechanistically, Kdm6b promotes the expression of the antiapoptotic gene Bcl2 and the cytotoxic genes Gzmb and Fasl in TCRαβ+CD8αα+ IELs through removal of the repressive H3K27me3 marker in enhancer and promoter regions .

What approaches can researchers use to study KDM6B's role in neuronal development using the FITC-conjugated antibody?

For investigating KDM6B in neuronal development:

• Developmental expression profiling:

  • Perform immunofluorescence on brain tissue sections across developmental stages

  • Use KDM6B Antibody, FITC conjugated with neuronal markers (NeuN, DCX, MAP2)

  • Quantify KDM6B expression changes during neuronal maturation

• In vitro neuronal differentiation models:

  • Apply KDM6B antibody staining in neural progenitor differentiation assays

  • Create time-course experiments to track KDM6B expression changes

  • Correlate with expression of mature neuronal gene programs

• Single-cell resolution techniques:

  • Combine flow cytometry using KDM6B Antibody, FITC conjugated with cell sorting

  • Perform RNA-seq on KDM6B-high versus KDM6B-low neuronal populations

  • Use fluorescent in situ hybridization combined with KDM6B immunofluorescence to correlate protein with mRNA expression

• Functional manipulation approaches:

  • Knockdown or overexpress KDM6B in neuronal cultures

  • Assess changes in neuronal morphology, synaptic connections, and electrophysiological properties

  • Compare epigenetic landscape (H3K27me3 distribution) between experimental conditions

Research insight: KDM6B has been implicated as a regulator of neuronal maturation beyond its previously established functions at early stages of neuronal differentiation. This suggests its continued importance throughout the neuronal development process .

How can researchers optimize KDM6B Antibody, FITC conjugated for multi-parameter flow cytometry in rare cell populations?

For optimizing KDM6B antibody use in multi-parameter flow cytometry:

• Panel design considerations:

  • Place KDM6B-FITC in appropriate channel based on expression level (FITC is medium brightness)

  • Avoid fluorophore combinations with significant spectral overlap with FITC

  • Include viability dye compatible with fixation/permeabilization required for KDM6B staining

  • Incorporate key surface markers for identifying cell subsets before fixation/permeabilization

• Rare cell detection protocol:

  • Start with enrichment steps (magnetic bead sorting, density gradient)

  • Increase event count (collect >500,000 events)

  • Implement hierarchical gating strategy to focus on populations of interest

  • Use fluorescence minus one (FMO) controls for precise gating

• Fixation and permeabilization optimization:

  • Test multiple fixation/permeabilization buffers to maximize nuclear antigen detection

  • Determine optimal fixation duration (typically 10-20 minutes)

  • Consider mild fixation for surface markers followed by stronger fixation/permeabilization for KDM6B

• Signal amplification strategies:

  • Use primary KDM6B antibody followed by FITC-conjugated secondary for signal enhancement

  • Consider tyramide signal amplification for very low abundance detection

  • Optimize antibody concentration through titration experiments

Example staining protocol for hematopoietic stem/progenitor cells:

• Stain fresh bone marrow cells with surface markers (CD34, CD38, lineage cocktail)

• Fix with 2% paraformaldehyde (15 minutes, 4°C)

• Permeabilize with 0.1% Triton X-100 (10 minutes, room temperature)

• Block with 2% normal goat serum (30 minutes, room temperature)

• Stain with KDM6B Antibody, FITC conjugated (optimal dilution, overnight at 4°C)

• Wash and analyze by flow cytometry

What are the best approaches for quantitative analysis of KDM6B expression using the FITC-conjugated antibody?

For quantitative analysis of KDM6B expression:

• Flow cytometry quantification:

  • Use calibration beads with known fluorophore molecules (MESF beads)

  • Create a standard curve of fluorescence intensity

  • Report KDM6B expression as molecules of equivalent soluble fluorochrome (MESF)

  • Include quantitative flow cytometry controls in each experiment

• Image-based quantification:

  • Establish consistent acquisition parameters (exposure time, gain)

  • Use reference standards in each imaging session

  • Implement automated analysis with nuclear segmentation

  • Measure parameters including:

    • Mean nuclear fluorescence intensity

    • Integrated density (sum of pixel values)

    • Nuclear/cytoplasmic ratio

• Western blot quantification:

  • Include protein loading standard curve

  • Use fluorescence-based detection systems for wider dynamic range

  • Normalize KDM6B signal to appropriate loading controls

  • Implement densitometry with standard software (ImageJ, Image Studio)

• Statistical considerations for quantitative analysis:

  • Determine coefficient of variation across technical replicates

  • Establish minimum detectable difference for power calculations

  • Use appropriate statistical tests based on data distribution

  • Consider batch effects in multi-experiment comparisons

How can researchers integrate KDM6B protein analysis with functional epigenetic assays?

To integrate KDM6B protein analysis with functional epigenetic assays:

• Sequential ChIP-western blot approach:

  • Perform chromatin immunoprecipitation (ChIP) with KDM6B antibody

  • Use half of the precipitated material for sequencing (ChIP-seq)

  • Use the other half for western blot with KDM6B Antibody, FITC conjugated

  • Correlate ChIP-seq peak intensity with western blot signal strength

• Combined KDM6B and histone modification analysis:

  • Perform multi-color immunofluorescence with:

    • KDM6B Antibody, FITC conjugated

    • Antibodies against H3K27me3 (target of KDM6B)

    • Antibodies against H3K4me3 (active transcription mark)

  • Quantify pixel-by-pixel correlation between modifications

  • Create colocalization maps for nuclear distribution

• Functional enzyme activity assays:

  • Immunoprecipitate KDM6B using compatible antibodies

  • Perform in vitro demethylase assays with H3K27me3 peptides

  • Correlate enzyme activity with protein expression levels

  • Test effects of inhibitors (e.g., GSK-J4) on KDM6B activity

• Gene expression correlation studies:

  • Sort cells based on KDM6B-FITC signal intensity

  • Perform RNA-seq on KDM6B-high versus KDM6B-low populations

  • Identify differentially expressed genes

  • Validate regulation mechanism through ChIP at candidate loci

Research insight: In functional studies, KDM6B has been shown to promote gene expression through dual mechanisms: removing the repressive H3K27me3 mark and positively regulating the activating H3K4me3 modification through interaction with Set1/MLL methyltransferase complexes .

How might KDM6B Antibody, FITC conjugated be used in investigating the role of KDM6B in inflammatory diseases?

For studying KDM6B in inflammatory diseases:

• Inflammatory cell population analysis:

  • Develop multi-parameter flow cytometry panels combining:

    • KDM6B Antibody, FITC conjugated

    • Inflammatory cell markers (CD14, CD16 for monocytes; CD3, CD4 for T cells)

    • Activation markers (CD80, CD86, HLA-DR)

  • Compare KDM6B expression in cells from inflamed versus healthy tissues

• Tissue inflammation studies:

  • Perform immunofluorescence on tissue sections from inflammatory disease models

  • Co-stain with KDM6B Antibody, FITC conjugated and:

    • Inflammatory cytokine markers (TNF-α, IL-6)

    • Cell-specific markers

    • Signaling pathway components (NF-κB, STAT proteins)

• Response to inflammatory stimuli:

  • Monitor KDM6B expression changes following TLR ligand exposure (e.g., LPS)

  • Track kinetics of KDM6B upregulation during inflammatory response

  • Correlate with production of inflammatory mediators (cytokines, chemokines)

• Therapeutic intervention assessment:

  • Test effects of anti-inflammatory drugs on KDM6B expression

  • Evaluate KDM6B inhibitors (e.g., GSK-J4) in inflammatory disease models

  • Monitor both KDM6B protein levels and downstream inflammatory gene expression

Research insight: KDM6B plays a central role in inflammatory responses by participating in macrophage differentiation during inflammation through regulating gene expression. Studies have shown that KDM6B overexpression can lead to increased levels of inflammatory cytokines like tumor necrosis factor and CXCL2 (MIP-2) in peripheral blood .

What are potential applications of KDM6B Antibody, FITC conjugated in cancer immunotherapy research?

For cancer immunotherapy research applications:

• Tumor-infiltrating lymphocyte (TIL) analysis:

  • Use flow cytometry with KDM6B Antibody, FITC conjugated to assess expression in different TIL subsets

  • Compare KDM6B levels in:

    • Effector vs. exhausted T cells

    • Conventional vs. regulatory T cells

    • Tumor-associated macrophages with M1 vs. M2 phenotypes

• Immune checkpoint interaction studies:

  • Evaluate KDM6B expression in relation to immune checkpoint molecules (PD-1, CTLA-4)

  • Analyze how checkpoint blockade therapy affects KDM6B expression

  • Investigate KDM6B's epigenetic regulation of checkpoint genes

• Epigenetic immunotherapy combination approaches:

  • Test KDM6B inhibitors in combination with checkpoint inhibitors

  • Monitor changes in tumor microenvironment immune profiles

  • Assess effects on cytotoxic function of immune cells

• Biomarker development strategy:

  • Evaluate KDM6B expression as a potential predictive biomarker for immunotherapy response

  • Correlate baseline KDM6B levels with treatment outcomes

  • Develop standardized flow cytometry protocols for clinical sample analysis

Research insight: Studies in mouse models have shown that Kdm6b promotes the expression of cytotoxic genes like Gzmb and Fasl in intestinal intraepithelial lymphocytes through removal of repressive H3K27me3 marks . This mechanistic insight suggests KDM6B may play important roles in regulating cytotoxic functions of tumor-infiltrating lymphocytes as well.

How can researchers use KDM6B Antibody, FITC conjugated in single-cell epigenomic analyses?

For integrating KDM6B antibody in single-cell epigenomic approaches:

• Advanced single-cell protein-genomic integration:

  • Implement CITE-seq or REAP-seq approaches incorporating KDM6B Antibody, FITC conjugated

  • Sort cells based on KDM6B-FITC levels for downstream single-cell ATAC-seq

  • Correlate KDM6B protein levels with chromatin accessibility patterns

• Spatial epigenomics with KDM6B detection:

  • Perform in situ KDM6B protein detection with FITC-conjugated antibody

  • Follow with in situ Hi-C or chromatin conformation capture techniques

  • Create spatial maps of nuclear organization in relation to KDM6B expression

• Computational integration methods:

  • Develop algorithms to integrate KDM6B protein levels with:

    • Single-cell transcriptome data

    • Chromatin accessibility profiles

    • DNA methylation patterns

  • Create multi-modal data visualization approaches

• Live-cell epigenetic dynamics:

  • Implement partial cell permeabilization techniques for nuclear KDM6B detection

  • Combine with live-cell reporters for chromatin dynamics

  • Track temporal changes in KDM6B localization and activity

Technical workflow example:

• Incubate cells with KDM6B Antibody, FITC conjugated

• Sort cells by FACS into KDM6B-high and KDM6B-low populations

• Process sorted populations for single-cell ATAC-seq or ChIP-seq

• Analyze differential chromatin accessibility at KDM6B target genes

• Integrate with single-cell RNA-seq data from matched populations

This integrated approach allows researchers to connect KDM6B protein levels directly to chromatin state and transcriptional output at single-cell resolution, providing unprecedented insights into the epigenetic mechanisms of cellular heterogeneity.