Fig. 2.

DEPTOR knockout promotes cell proliferation and clonogenic survival and the activation of S6K1 and AKT in prostate cancer cells. a DEPTOR knockout promotes the proliferation of prostate cancer cells. DEPTOR-knockout DU145 and 22RV1 cells were generated via CRISPR/Cas9 technology. Cells were seeded in 96-well plates in triplicate and then subjected to an ATPlite-based cell proliferation assay. Cell proliferation is expressed as the fold change compared with that at day 1. The mean ± SEM are shown from three independent experiments, n = 3; **p < 0.01; ***p < 0.001. b DEPTOR knockout promotes clonogenic survival of prostate cancer cells. Cells were seeded in 60-mm dishes at 500 cells per dish in triplicate and were incubated for 7–14 days, followed by staining (left) and colony counting (right). The mean ± SEM are shown from three independent experiments; n = 3; ***p < 0.001. c DEPTOR knockout causes increased phosphorylation of the mTORC1 and mTORC2 downstream effectors, S6K1 and AKT, respectively. Cells were harvested for western blotting using the indicated antibodies. The band density was quantified and expressed as the relative gray value (compared with the control), by arbitrarily setting the control value as 1