Journal: bioRxiv

Article Title: Chromatin modifiers KMT2D, BAF, and p300 are required for de novo binding of transcription factors on enhancers

doi: 10.64898/2026.01.29.702555

Figure Lengend Snippet: (A) Schematic of the 4-OHT-inducible MyoD-ER system. (B) MyoD expression in preadipocytes and C2C12 myoblasts was determined using RNA-Seq (n = 1). RPKM values indicate gene expression levels. (C) Western blot (WB) analysis of nuclear extracts from preadipocytes expressing MyoD-ER-T7 and treated with 4-OHT. Antibodies used were indicated on the right. BRG1 was used as a loading control. (D-I) MyoD-ER-T7 expressing preadipocytes were treated with 4-OHT for 1 hour (h), followed by CUT&RUN analysis. (D) Bar chart showing ARID1A (an exclusive subunit of BAF), KMT2D, and p300 binding status on induced MyoD sites. (E) Box plots displaying the normalized MyoD read counts in subgroups defined in (D). (F) Bar chart showing ARID1A (BAF), KMT2D, and p300 binding on 38,732 MyoD + enhancers defined in (D) prior to 4-OHT treatment. (G-H) Box plots showing the normalized MyoD read counts (G) and HOMER de novo motif analysis (H) on BAF-KMT2D-p300 prebound or de novo sites defined in (F). Statistical significance was determined using a two-sided, unpaired Mann Whitney test. (I) Genome browser view of MyoD binding sites around Maged1 and Cap2 loci.

Article Snippet: BRG1 Inhibitor BRM014 (#HY-119374) and 5Ph-IAA (#HY-134653) were from MedChemExpress (MCE) and used at 1μM.

Techniques: Expressing, RNA Sequencing, Gene Expression, Western Blot, Control, Binding Assay, MANN-WHITNEY

Journal: bioRxiv

Article Title: Chromatin modifiers KMT2D, BAF, and p300 are required for de novo binding of transcription factors on enhancers

doi: 10.64898/2026.01.29.702555

Figure Lengend Snippet: (A) Sanger sequencing results verifying the integration of 132-bp AID sequence after the start codon (ATG) of Kmt2d . (B) Breeding scheme for generating Kmt2c f/f ; Kmt2d AID/AID ; Cre-ER mice. (C) Diagram illustrating the workflow to generate Kmt2c -/- ; Kmt2d AID/AID preadipocytes expressing OsTIR1-F74G-Myc and MyoD-ER-T7. (D) WB of nuclear extracts confirming KMT2C knockout. Antibodies used were indicated on the right. BRG1 was a loading control. (E) WB of nuclear extracts confirming the depletion of KMT2D in Kmt2d AID/AID ; MyoD-ER-T7 preadipocytes upon 5Ph-IAA treatment for various time durations. Antibodies used were indicated on the right. RbBP5 was a loading control.

Article Snippet: BRG1 Inhibitor BRM014 (#HY-119374) and 5Ph-IAA (#HY-134653) were from MedChemExpress (MCE) and used at 1μM.

Techniques: Sequencing, Expressing, Knock-Out, Control

Journal: bioRxiv

Article Title: Chromatin modifiers KMT2D, BAF, and p300 are required for de novo binding of transcription factors on enhancers

doi: 10.64898/2026.01.29.702555

Figure Lengend Snippet: Kmt2d AID/AID ; MyoD-ER-T7 preadipocytes were pretreated with BRG1 inhibitor BRM014 (BRG1i) for 1h, and then 4-OHT was added for 1h to induce MyoD nuclear translocation. Cells were harvested for WB, CUT&RUN, and ATAC-seq. (A) WB of nuclear extracts for KMT2D, p300, and BAF subunits BRG1 and ARID1A. (B) Pie chart illustrating BAF binding status on 38,732 MyoD + enhancers. (C) Heat maps for CUT&RUN of ARID1A (BAF), T7 (MyoD), KMT2D, and p300 on BAF pre-bound and de novo BAF binding sites. (D-E) Chromatin accessibility determined by ATAC-seq signals on MyoD + enhancers. Chromatin accessibility status on BAF prebound sites (D) or de novo BAF binding sites (E) is shown in pie charts ( upper panels ). Average profiles of normalized ATAC-seq reads on constitutively open and MyoD-dependent opening sites are shown in lower panels .

Article Snippet: BRG1 Inhibitor BRM014 (#HY-119374) and 5Ph-IAA (#HY-134653) were from MedChemExpress (MCE) and used at 1μM.

Techniques: Translocation Assay, Binding Assay