Fig. 2.

HULLK is an AR-regulated lncRNA. a HULLK is positively regulated by androgen, n = 3. LNCaP and C42 cells were transduced with two independent shRNAs to LCK and treated with 0-1 nM R1881. RNA was collected on Day 4 and LCK transcript levels were determined using LCK primers targeting the 3’UTR. b Enzalutamide blocks the androgen-induced expression of HULLK, n = 3. LNCaP and C42 cells were seeded for 48 h, and then, treated with 1 nM R1881 in the presence or absence of 10 μM enzalutamide. RNA was collected 24 h after treatment and LCK transcript levels were determined by qPCR using LCK primers targeting the 3’UTR. c HULLK is not upregulated in AR-null PCa cells, n = 3. C4–2, DU145, and PC3 cells were seeded in CSS media for 48 h, and then, treated with 1 nM R1881 for 16 h. RNA was collected and LCK transcript levels were determined using LCK primers targeting the 3’UTR. d Inhibition of p300 blocks the increase in HULLK expression induced by hormone, n = 3. e Brd4 inhibition reduces the hormone induction of HULLK, n = 3. LNCaP, C42, or Jurkat cells were seeded for 48 h in the appropriate growth media, and then, treated with 1 nM R1881 in the presence or absence of 0.3 μM A-485 or 0.1 μM JQ1. RNA was collected 24 h after treatment and LCK transcript levels were determined by qPCR using LCK primers targeting exon 13 or 3’UTR. PSA and TMPRSS2 transcripts were determined to verify the efficacy of p300 and Brd4 inhibition