Overexpression of EP4 with cAMP-PKA signal activation promoted the castration-resistant progression of LNCaP cells through AR activation. Overexpression.

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Supplemental Figure 1 a % pulldown from input LNCaP/scrambled-siRNA Figure 1 A, ChIP analysis of AR and PHB binding to the PSA promoter in the LNCaP/scrambled-siRNA.

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Overexpression of EP4 with cAMP-PKA signal activation promoted the castration-resistant progression of LNCaP cells through AR activation. Overexpression of EP4 with cAMP-PKA signal activation promoted the castration-resistant progression of LNCaP cells through AR activation. A, EP4 protein expression detected with Western blotting in LNCaP-EP4(A) and LNCaP-EP4(B) compared with LNCaP-mock (top), intracellular cAMP concentrations, PSA expression, and cell proliferation ratio versus day 0 in androgen-depleted conditions (CSFBS) for 6 d (bottom). *, P < 0.05; **, P < 0.005 versus LNCaP-mock. B, AR expression attenuated using a stealth RNAi system detected with Western blotting (top), PSA expression levels on day 4 (middle), and cell proliferation ratio on days 2 and 4 versus day 0 (bottom) in CSFBS. *, P < 0.05; **, P < 0.005 versus LNCaP-mock. C, relative luciferase activities of PSAp-Luc in LNCaP cells with transfection of pcDNA3.1-mock and pcDNA3.1-EP4 in CSFBS and 5α-dihydrotestosterone (DHT) stimulation, respectively. *, P < 0.05; **, P < 0.005 versus control (left). PSA expression of LNCaP cells in CSFBS for 6 d under the administration of forskolin, dbcAMP, and H-89. *, P < 0.05; **, P < 0.005 versus preadministration (right). D, the sequential changes in xenograft tumor volume after the castration of mice (1 × 107 cells, n = 5 each). *, P < 0.05. Naoki Terada et al. Cancer Res 2010;70:1606-1615 ©2010 by American Association for Cancer Research