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. 2020 Nov 12;11(11):976. doi: 10.1038/s41419-020-03185-3

Fig. 4. p53-mediated DEPTOR expression inhibits cell proliferation and survival via AKT inactivation.

Fig. 4

a Deletion of p53-binding site C on the DEPTOR promoter activates AKT signaling. Cells were serum-starved for 24 h, followed by serum supplementation and were then harvested at the indicated time points for IB with the indicated Abs. The band density was quantified and expressed as the ratio of p-AKT:t-AKT and the ratio of p-S6K1:t-S6K1 at various time points. b–c Silencing of AKT abrogates the induction of cell proliferation and survival by the deletion of site C. Cells with or without site C were transfected with the indicated siRNAs for 48 h, followed by ATPlite-based cell proliferation assay (b, left), IB with the indicated Abs (b, right), or clonogenic survival assay c. Cell proliferation is expressed as the fold change compared with day 1 (mean ± S.E.M, n = 3; *p < 0.05). Cell survival is expressed as the rate of colony formation (c, right) (mean ± S.E.M, n = 3; *p < 0.05, ***p < 0.001). d AKT inactivation abrogates the induction of cell proliferation by the deletion of site C. Cells with or without site C were seeded in triplicate in 96-well plates and then treated with MK-2206 or left untreated, followed by ATPlite-based cell proliferation assay. Cell proliferation is expressed as the fold change compared with day 1 (mean ± S.E.M, n = 3; **p < 0.01, ***p < 0.001).