Effects of specific kinase inhibitors on select signaling pathways and apoptosis in LNCaP and LNCaPH cells. LNCaP and LNCaPH cells were pretreated with 25 μM LY294002, 20 μM U0126, and 10 μM SP600125 alone or the combination of LY294002 and U0126 for 48 h. Control cells were grown in the presence of DMSO. A, note the percent of apoptotic cells with sub-G1 DNA content. Values represent the mean ± SE of three independent experiments. Asterisk indicates P < 0.05 compared with LNCaP cells. B, immunoblot analysis was performed using antibodies that specifically recognize total and phosphorylated Akt, ERK, JNK, and AR. Blots were stripped and reprobed with an antibody against β-tubulin.