Fig. 5

GATA3 promotes, but is not essential for, expression of shared AR and GATA3 target genes. A Representative immunofluorescence images showing AR silencing in T-47D cells. Scale bar = 30 μm. B GATA3 ChIP-PCR at AR/GATA3 co-occupied loci associated with DHT-regulated AR target genes following AR silencing by two different siRNAs in T-47D cells (upper panel) and MDA-MB-453 cells (lower panel), treated in vitro with Veh (EtOH) or DHT (10 nM). Data was analyzed by a two-way ANOVA. Post hoc analyses were performed using Tukey’s multiple comparisons test, where SEC14L2, P < 0.0001 for Veh versus DHT in siControl only; ZBTB16, P < 0.0001 and P < 0.01 for Veh versus DHT in siControl and siAR-1, respectively. Analysis of GATA3 chromatin binding at a locus associated with the C-FOC gene was included as a negative control for androgen-unresponsive GATA3 binding. C AR ChIP-PCR at AR/GATA3 co-occupied loci associated with DHT-regulated genes following GATA3 knockdown in T-47D cells (upper panel) and MDA-MB-453 cells (lower panel), treated in vitro with Veh (EtOH) or DHT (10 nM). Data was analyzed by a two-way ANOVA. Post hoc analyses were performed using Tukey’s multiple comparisons test, where SEC14L2, P < 0.0001, and P < 0.01 for Veh versus DHT in siControl and siGATA3, respectively, and P < 0.0001 for siControl + DHT versus siAR + DHT; ZBTB16, P < 0.0001 for Veh versus DHT in siControl and siGATA3, and P < 0.0001 for siControl + DHT versus siGATA3 + DHT. Analysis of AR chromatin binding at a locus associated with the FKBP5 gene was included as a control for GATA3-independent AR chromatin binding. Data is shown as mean ± SEM of 3 biological replicates from consecutive passages of cells