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. 2021 Jul 24;40(37):5590–5599. doi: 10.1038/s41388-021-01911-5

Fig. 3. mTOR inhibitors induce cell cycle arrest and apoptosis in CR melanoma cells.

Fig. 3

A Relative survival of WM164-CR, A2058-CR, UACC903-CR, and WM9-CR cells treated with increasing concentrations of Rapamycin (Rapa, Max = 100 µM), NVP-BEZ235 (BEZ, Max = 100 µM), PLX4720 (PLX, Max = 100 µM) and PD0325901 (PD, Max = 10 µM) for 72 h. B Heatmaps of indicated pathway scores from RPPA analysis of A2058-CR and UACC903-CR cells after treatment with PLX (2.5 µM) + PD (0.25 µM) in the absence or presence of Rapa (100 nM) and BEZ (1 µM) for 24 h. C The expression of cell cycle and apoptosis -related proteins was analyzed by western blotting in CR cells treated with indicated inhibitors (PLX, 2.5 µM; PD, 0.25 µM; Rapa, 100 nM; BEZ, 1 µM) for 72 h. D Quantification of EdU incorporation assays performed to detect the proliferation of WM164-CR, UACC903-CR, and A2058-CR melanoma cells treated with Rapamycin (Rapa, 100 nM) and NVP-BEZ235 (BEZ, 1 µM) in the presence of PLX (2.5 µM) and PD (0.25 µM) for 24 h. E Flow cytometry analysis of cell cycle stages of cells in (D). F Quantification of early and late apoptotic cell population treated with Rapamycin and NVP-BEZ235 in the presence of BRAF and MEK inhibitors by the Annexin V/7-AAD double staining. Data are presented as mean ± SD (n = 3). P values (vs. PLX + PD treatment group) are from a two-sided unpaired Student’s t-test.