Figure 2.
Figure 2. BRD4 binds regulatory regions in CTCL patients. (A) ChIP analysis of BRD4 binding to active regions of the genome in CD4+ T cells from a normal donor (left), CTCL patient (middle), and CTCL patient cells treated with 100 nM JQ1 in vitro for 24 hours. (B-D) ChIP analysis of BRD4 binding to (B) promoter regions, (C) distal active regions, and (D) super-enhancer regions. (E) Average signal intensity plot of BRD4 binding to active regions of the genome in CD4+ T cells from a normal donor, CTCL patient, and CTCL patient cells treated with 100 nM JQ1 in vitro for 24 hours. (F-H) Average signal intensity plot of BRD4 binding to (F) promoter regions, (G) distal active regions, and (H) super-enhancer regions. (I) Pearson correlation map demonstrating correlation coefficients of BRD4 binding across all genomic regulatory regions between normal donor, CTCL patient, and patient treated with JQ1. (J) MTS assay showing viability of CTCL-derived cell lines exposed to increasing doses of JQ1 from 0.25 µM to 20 µM. All cell lines demonstrate <50% cell viability at high doses of JQ1. Fifty percent effective concentration (EC50) values are presented as a table. (K) Cell cycle analysis of CTCL-derived cell lines exposed to increasing doses of JQ1 from 0.5 µM to 10 µM demonstrating the percentage of cells in sub-G0/G1 phase. Data are presented as mean ± SEM unless otherwise specified.

BRD4 binds regulatory regions in CTCL patients. (A) ChIP analysis of BRD4 binding to active regions of the genome in CD4+ T cells from a normal donor (left), CTCL patient (middle), and CTCL patient cells treated with 100 nM JQ1 in vitro for 24 hours. (B-D) ChIP analysis of BRD4 binding to (B) promoter regions, (C) distal active regions, and (D) super-enhancer regions. (E) Average signal intensity plot of BRD4 binding to active regions of the genome in CD4+ T cells from a normal donor, CTCL patient, and CTCL patient cells treated with 100 nM JQ1 in vitro for 24 hours. (F-H) Average signal intensity plot of BRD4 binding to (F) promoter regions, (G) distal active regions, and (H) super-enhancer regions. (I) Pearson correlation map demonstrating correlation coefficients of BRD4 binding across all genomic regulatory regions between normal donor, CTCL patient, and patient treated with JQ1. (J) MTS assay showing viability of CTCL-derived cell lines exposed to increasing doses of JQ1 from 0.25 µM to 20 µM. All cell lines demonstrate <50% cell viability at high doses of JQ1. Fifty percent effective concentration (EC50) values are presented as a table. (K) Cell cycle analysis of CTCL-derived cell lines exposed to increasing doses of JQ1 from 0.5 µM to 10 µM demonstrating the percentage of cells in sub-G0/G1 phase. Data are presented as mean ± SEM unless otherwise specified.

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