Figure 6

INPP4B regulates IL-8 gene expression through PKC in LNCaP cells. (A-B) Regulation of IL-8 by INPP4B in LNCaP cells was evaluated 48 hours after INPP4B knockdown. RNA was extracted and analyzed for expression of INPP4B (A) and IL-8 (B) by quantitative RT-PCR and normalized to 18S. (C) LNCaP cells cultured in 10% CSS medium were treated with 5 μM AZD5363 or 10 μM LY294002 for 24 hours. RNA was extracted and analyzed for expression of IL-8 (C) by quantitative RT-PCR and normalized to 18S. (D) Inhibition of PI3K and Akt by LY294002 and AZD5363 was confirmed by western blot analysis of phosphorylation of ribosomal protein S6 following treatment with 0.5 or 5 μM AZD5363 and 10 μM LY294002. (E) LNCaP cells cultured in 10% CSS media were treated with BIM-I (BIM) or PMA. RNA was extracted and analyzed for expression of IL-8 by quantitative RT-PCR. (F) LNCaP cells were treated with DMSO (V) or 2 μM BIM-I (B), 250 nM PMA (P) or BIM-I plus PMA (P/B) to inhibit or activate PKC signaling. Proteins were analyzed for phospho-S6 and total S6 protein levels. Gene expression data are presented as means ± SEM. * P < 0.05, **P < 0.01, *** P < 0.001.