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. 2020 Dec 21;2(4):zcaa039. doi: 10.1093/narcan/zcaa039

Figure 6.

Figure 6.

CDC25B modulation of metformin cytotoxicity via PP2A-AMPK. (A) BT549 and HS578T cells were transfected with EV, CDC25B or PP2A-C plasmids, negative siRNA or siAMPKα1 and α2, and then treated with increasing doses of metformin for 72 h, and cell survival was then determined. The x-axis indicates drug dose, and the y-axis indicates the survival fraction after metformin exposure. Knockdown or overexpression efficiency is shown in mRNA levels. (B) CDC25B knockdown BT549 and HS578T were transfected with siPP2A, and then treated with increasing doses of metformin for 72 h, and cell survival was then determined. The x-axis indicates drug dose, and the y-axis indicates the survival fraction after metformin exposure. Knockdown efficiency is shown in mRNA levels. (C) CDC25B knockout BT549 and stable knockdown HS578T cells transfected with EV, full length CDC25B (FL), CDC25B N-terminal domain (1–375 aa) (NTD), or CDC25B C-terminal domain (374–580 aa) (CTD), were treated with increasing doses of metformin for 72 h and cell survival was then determined. The x-axis indicates drug dose, and the y-axis indicates the survival fraction after metformin exposure. (D) CDC25B knockout BT549 and stable knockdown HS578T cells transfected with EV, WT CDC25B (WT), CDC25B S353A, CDC25B S353E, or CDC25B C488S constructs, were treated with increasing doses of metformin for 72 h and cell survival was then determined. The x-axis indicates drug dose, and the y-axis indicates the survival fraction after metformin exposure. (E) Cells were transfected with indicated siRNA, and then treated with increasing doses of metformin for 72 h, and cell survival was then determined. The x-axis indicates drug dose, and the y-axis indicates the survival fraction after metformin exposure. Data information: all data presented are in the format of mean ± SEM of N = 3 independent experiments with three biological replicates for each experiment. Statistically significant differences were determined using the two-way ANOVA plus Tukey (**P < 0.01), ns represents not significant.