INPP4B regulates IL-8 and PAK6 through PKC signaling in PC-3 cells. (A-B) PC-3 cells cultured in regular growth medium were treated with DMSO (vehicle), 2 μM BIM-I (BIM), or 250 nM PMA to inhibit or activate PKC signaling respectively. RNA was extracted and analyzed for expression of IL-8 (A) and PAK6 (B) by quantitative RT-PCR and normalized to 18S. (C) PC-3 Tet-On Neg and #14 clones were cultured for 2 days ± doxycycline and treated with 250 nM PMA for 4.5 hours prior to RNA extraction and gene analysis for IL-8. (D) PC-3 Tet-On #14 cells were cultured without Dox (Con), or with Dox for 2 days prior to protein extraction. Lysates were analyzed for phospho-PKCβII S660, phospho-PKCζ T410, FLAG-INPP4B, and tubulin. (E) PC-3 Tet-On #14 cells were cultured with doxycycline for the indication time periods prior to protein extraction. Lysates were analyzed for BIRC5, FLAG-INPP4B, and tubulin protein levels by Western blot analysis. Data in A, B, and C are presented as means ± SEM.