However, MAP3K11 knockdown increased transcription of SGK in C4-2B cells 2 hours after the addition or hormone. Knockdown of DGKD decreased transcription of TMPRSS2 in C4-2B cells and of SGK in LNCaP cells at 24 hours. There was no change in TMPRSS2 or SGK transcription in either cell line as a result of the knockdown of ICK or PSKH1. CIT knockdown has disparate effects on AR transcription. Interestingly, the most consistent effect on AR transcription was from GALK2 knockdown, which caused an increase in SGK at 24 hours after hormone addition in both cell lines and at 2 hours in C4-2B cells. However, examining only TMPRSS2 and SGK as representative AR target genes may create selection bias; therefore, we expanded our analysis of ARregulated genes. We analyzed 14 additional genes, including the AR-activated genes PSA, FKBP51, ORM1, STAG, Nkx3.1, FASN, AQP3, KLK2, and UGT2B; the AR-repressed genes DKK and FST; and the castration-resistant prostate cancer AR-regulated genes CDC20, CDK1, and UBE2C. Transcription in LNCaP cells following MAP3K11 knockdown was examined at 24 hours after androgen treatment. As expected, androgen RG7204 918504-65-1 induced or repressed transcription of all AR target genes in pLKO control cells. The effect of MAP3K11 knockdown on AR transcription is target dependent. Androgen-stimulated transcription of TMPRSS2, SGK, and ORM1 was reduced in response to MAP3K11 knockdown. The inverse was true for the androgen-repressed genes DKK and FST. MAP3K11 knockdown stimulated gene expression and diminished the amount of androgen-induced repression. PSA, FKBP51, STAG, Nkx3.1, FASN, AQP3, KLK2, and UGT2B did not change in response to MAP3K11 knockdown. Insofar as this subset of AR target genes represents the AR transcriptome, these data suggest that the inhibition of cell growth in response to MAP3K11 kinase knockdown may be due to regulation of AR transcriptional activity on a subset of AR-regulated genes. The AR selectively regulates cell cycle regulated genes such as CDC20, CDK1, and UBE2C to promote cell growth. We found that MAP3K11 knockdown led to a slight, but statistically significant reduction in transcription of these M-phase AR-regulated genes. Previously, we demonstrated that stress kinases could regulate AR Ser 650 phosphorylation. Thus, to further explore the mechanism of MAP3K11 regulation of prostate cancer cell growth and AR transcription, we tested if MAP3K11 knockdown regulated AR Ser 650 phosphorylation since MAP3K11 is an upstream regulator of JNK activity. PMA induced AR Ser 650 phosphorylation in LNCaP and C4-2B cells more than 3 fold. In these experiments, both shRNAs decreased MAP3K11 protein expression, with MAP3K11 shRNA-1 decreasing expression to a greater extent than MAP3K11 shRNA-2. Similar alterations in c-Jun phosphorylation were observed, suggesting that the MAP3K11 shRNAs inhibited PMA-induced stress kinase signaling. Under these conditions, AR Ser 650 phosphorylation was decreased in both LNCaP and C4-2B cells. A parallel decrease in total AR was observed.