A, Immunoblot of phospho-Smad3 (pSmad3) and Smad3 in DU145
cells treated with either 0.01% DMSO or 100 nM MZ1 for 21 hours, serum starved
for 2 hours under the same conditions and then dosed with 5 ng/mL TGFß
for the indicated time. The ratio of pSmad3:Smad3 illustrates Smad3 activation
and quantifications are relative to control. Blots shown are representative of
two independent experiments.
B, DU 145 cells treated with either 400 nM of (–)JQ1
or (+)JQ1 for 24 hours and then harvested for ChIP. BRD4 interacts with the
SNAI1 and SNAI2 promoters and is displaced
upon exposure to (+)JQ1. Results from two independent experiments are shown.
Statistical analyses were performed using the Student’s t test.
Significant differences: ns, nonsignificant, P > 0.05;
*.
C, Publicly available ChIP-seq datasets performed on DHT or
DHT + JQ1-treated VCaP cells were analyzed for the binding of BRD4 and RNA Pol
II at the SNAI1 and SNAI2 locus. The analyzed
datasets are accessible on the GEO platform (GSE55062 (16)). Data visualization was performed using
Integrative Genomics Viewer (IGV).