Fig 4. Dose-dependent effects of GPR158 in LNCaP cells.

(A and B) LNCaP cells stably infected with lentivirus constructs containing GPR158 driven by a doxycycline inducible promoter were treated with low dose (100 ng/mL) or high dose (500 ng/mL) doxycycline concentration for 3-days. (A) The cell lysates were subjected to western blotting for the confirmation of induction of GPR158, AR, PSA and beta-actin proteins. (B and C) After doxycycline treatment for 3-days at indicated dose (B) and indicated time-points at 100ng/mL for growth curve (C), the cells were trypsinized and counted using trypan blue dye in a hemocytometer chamber. The data represent normalized values with separate experiment performed to estimate the effect of doxycycline alone on parental LNCaP cell numbers under the same experimental conditions as above. (B and C) The results are indicative of three independent experiments, each performed in triplicate.