Fig. 3: PI3K family is crucial for Brusatol-mediated inhibition.

a IC50 assays of Brusatol were determined in the indicated cancer cells. All of the cells were treated for 72 h. b The indicated Brusatol-less sensitive or -more sensitive cells were treated with 100 nM Brusatol for indicated periods (0, 24 h, 48 h, 72 h), and cell viability was examined by detecting luminescence. Results are the mean ± standard error of the duplicates. ****p < 0.0001 shows the significant differences between the indicated groups. NS, not significant. c Brusatol-less sensitive or -more sensitive cells were treated with 100 nM Brusatol for different periods (0, 24 h, 48 h). Then cells were fixed and stained with PI. Flow cytometry assays were performed to determine cell cycle progression. The subG1 population in cells was labeled as the percentage. d The endogenous expression of PI3K/AKT signaling protein in these cell lines, including sets of Brusatol-less sensitive and -more sensitive cells, was detected with western blot. e The described cells were incubated with Brusatol (0, 50 nM, 100 nM) for different periods (0, 12 h, 24 h). Western blot analysis was performed to determine the expression of PI3K/AKT regulated signaling proteins in Brusatol-less sensitive cells set (HL-60, K562) and Brusatol-more sensitive cells set (Raji, SU-DHL-4).