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(A) Flow cytometry analysis of H3K27me3 and H3K36me3 intracellular staining in mesenchymal stem cells expressing mutant histones, y -axis showing normalized event count. (B) Immunoblot of H3K27me3 and H3K36me3 in acid-extracted histones from mesenchymal stem cells expressing individual mutant histones. (C) Linear fit of total histone H3 intracellular staining over DAPI staining in G2/M vs G1 cell population. Individual data points colored by the histone PTM stained in addition to total histone H3. (D) Coomassie staining of recombinantly purified Tn5 <t>after</t> <t>Ni-NTA</t> column purification (left) and ion-exchange column purification (right), highlighted with the pooled fractions concentrated for the final product. (E) Immunoblot of acetyl-lysine in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. (F) Contour density plot of ATAC-see signal (Alexa Fluor 488) vs DAPI signal (Pacific Blue) in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. Histogram of DAPI signal (shown right) used to gate cell population into G1, S, and G2/M. (G) Correlation plot of PottsMPNN prediction results and published nucleosome dimer exchange data.
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(A) Flow cytometry analysis of H3K27me3 and H3K36me3 intracellular staining in mesenchymal stem cells expressing mutant histones, y -axis showing normalized event count. (B) Immunoblot of H3K27me3 and H3K36me3 in acid-extracted histones from mesenchymal stem cells expressing individual mutant histones. (C) Linear fit of total histone H3 intracellular staining over DAPI staining in G2/M vs G1 cell population. Individual data points colored by the histone PTM stained in addition to total histone H3. (D) Coomassie staining of recombinantly purified Tn5 <t>after</t> <t>Ni-NTA</t> column purification (left) and ion-exchange column purification (right), highlighted with the pooled fractions concentrated for the final product. (E) Immunoblot of acetyl-lysine in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. (F) Contour density plot of ATAC-see signal (Alexa Fluor 488) vs DAPI signal (Pacific Blue) in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. Histogram of DAPI signal (shown right) used to gate cell population into G1, S, and G2/M. (G) Correlation plot of PottsMPNN prediction results and published nucleosome dimer exchange data.
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(A) Flow cytometry analysis of H3K27me3 and H3K36me3 intracellular staining in mesenchymal stem cells expressing mutant histones, y -axis showing normalized event count. (B) Immunoblot of H3K27me3 and H3K36me3 in acid-extracted histones from mesenchymal stem cells expressing individual mutant histones. (C) Linear fit of total histone H3 intracellular staining over DAPI staining in G2/M vs G1 cell population. Individual data points colored by the histone PTM stained in addition to total histone H3. (D) Coomassie staining of recombinantly purified Tn5 <t>after</t> <t>Ni-NTA</t> column purification (left) and ion-exchange column purification (right), highlighted with the pooled fractions concentrated for the final product. (E) Immunoblot of acetyl-lysine in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. (F) Contour density plot of ATAC-see signal (Alexa Fluor 488) vs DAPI signal (Pacific Blue) in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. Histogram of DAPI signal (shown right) used to gate cell population into G1, S, and G2/M. (G) Correlation plot of PottsMPNN prediction results and published nucleosome dimer exchange data.
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(A) Flow cytometry analysis of H3K27me3 and H3K36me3 intracellular staining in mesenchymal stem cells expressing mutant histones, y -axis showing normalized event count. (B) Immunoblot of H3K27me3 and H3K36me3 in acid-extracted histones from mesenchymal stem cells expressing individual mutant histones. (C) Linear fit of total histone H3 intracellular staining over DAPI staining in G2/M vs G1 cell population. Individual data points colored by the histone PTM stained in addition to total histone H3. (D) Coomassie staining of recombinantly purified Tn5 <t>after</t> <t>Ni-NTA</t> column purification (left) and ion-exchange column purification (right), highlighted with the pooled fractions concentrated for the final product. (E) Immunoblot of acetyl-lysine in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. (F) Contour density plot of ATAC-see signal (Alexa Fluor 488) vs DAPI signal (Pacific Blue) in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. Histogram of DAPI signal (shown right) used to gate cell population into G1, S, and G2/M. (G) Correlation plot of PottsMPNN prediction results and published nucleosome dimer exchange data.
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(A) Flow cytometry analysis of H3K27me3 and H3K36me3 intracellular staining in mesenchymal stem cells expressing mutant histones, y -axis showing normalized event count. (B) Immunoblot of H3K27me3 and H3K36me3 in acid-extracted histones from mesenchymal stem cells expressing individual mutant histones. (C) Linear fit of total histone H3 intracellular staining over DAPI staining in G2/M vs G1 cell population. Individual data points colored by the histone PTM stained in addition to total histone H3. (D) Coomassie staining of recombinantly purified Tn5 <t>after</t> <t>Ni-NTA</t> column purification (left) and ion-exchange column purification (right), highlighted with the pooled fractions concentrated for the final product. (E) Immunoblot of acetyl-lysine in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. (F) Contour density plot of ATAC-see signal (Alexa Fluor 488) vs DAPI signal (Pacific Blue) in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. Histogram of DAPI signal (shown right) used to gate cell population into G1, S, and G2/M. (G) Correlation plot of PottsMPNN prediction results and published nucleosome dimer exchange data.
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(A) Flow cytometry analysis of H3K27me3 and H3K36me3 intracellular staining in mesenchymal stem cells expressing mutant histones, y -axis showing normalized event count. (B) Immunoblot of H3K27me3 and H3K36me3 in acid-extracted histones from mesenchymal stem cells expressing individual mutant histones. (C) Linear fit of total histone H3 intracellular staining over DAPI staining in G2/M vs G1 cell population. Individual data points colored by the histone PTM stained in addition to total histone H3. (D) Coomassie staining of recombinantly purified Tn5 <t>after</t> <t>Ni-NTA</t> column purification (left) and ion-exchange column purification (right), highlighted with the pooled fractions concentrated for the final product. (E) Immunoblot of acetyl-lysine in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. (F) Contour density plot of ATAC-see signal (Alexa Fluor 488) vs DAPI signal (Pacific Blue) in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. Histogram of DAPI signal (shown right) used to gate cell population into G1, S, and G2/M. (G) Correlation plot of PottsMPNN prediction results and published nucleosome dimer exchange data.
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(A) Flow cytometry analysis of H3K27me3 and H3K36me3 intracellular staining in mesenchymal stem cells expressing mutant histones, y -axis showing normalized event count. (B) Immunoblot of H3K27me3 and H3K36me3 in acid-extracted histones from mesenchymal stem cells expressing individual mutant histones. (C) Linear fit of total histone H3 intracellular staining over DAPI staining in G2/M vs G1 cell population. Individual data points colored by the histone PTM stained in addition to total histone H3. (D) Coomassie staining of recombinantly purified Tn5 after Ni-NTA column purification (left) and ion-exchange column purification (right), highlighted with the pooled fractions concentrated for the final product. (E) Immunoblot of acetyl-lysine in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. (F) Contour density plot of ATAC-see signal (Alexa Fluor 488) vs DAPI signal (Pacific Blue) in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. Histogram of DAPI signal (shown right) used to gate cell population into G1, S, and G2/M. (G) Correlation plot of PottsMPNN prediction results and published nucleosome dimer exchange data.

Journal: bioRxiv

Article Title: Multiomic screening platform uncovers the impact of histone mutations on chromatin and cell fate

doi: 10.64898/2026.04.14.718582

Figure Lengend Snippet: (A) Flow cytometry analysis of H3K27me3 and H3K36me3 intracellular staining in mesenchymal stem cells expressing mutant histones, y -axis showing normalized event count. (B) Immunoblot of H3K27me3 and H3K36me3 in acid-extracted histones from mesenchymal stem cells expressing individual mutant histones. (C) Linear fit of total histone H3 intracellular staining over DAPI staining in G2/M vs G1 cell population. Individual data points colored by the histone PTM stained in addition to total histone H3. (D) Coomassie staining of recombinantly purified Tn5 after Ni-NTA column purification (left) and ion-exchange column purification (right), highlighted with the pooled fractions concentrated for the final product. (E) Immunoblot of acetyl-lysine in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. (F) Contour density plot of ATAC-see signal (Alexa Fluor 488) vs DAPI signal (Pacific Blue) in mesenchymal stem cells treated with DMSO or 10 nM HDACi panobinostat overnight. Histogram of DAPI signal (shown right) used to gate cell population into G1, S, and G2/M. (G) Correlation plot of PottsMPNN prediction results and published nucleosome dimer exchange data.

Article Snippet: The supernatant was incubated with 4 mL pre-equilibrated Ni-NTA agarose resin (Qiagen) for 4 hours at 4 °C with gentle rotation.

Techniques: Flow Cytometry, Staining, Expressing, Mutagenesis, Western Blot, Purification