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

Figure 2.

Figure 2.

CDC25B interacts with PP2A. (A) HEK-293T cells were transfected with EV or Flag-CDC25B plasmids for 24 h then treated with 10 mM metformin for 48 h. Cell lysates were subjected to immunoprecipitation with anti-Flag antibody. The immunoprecipitates were blotted with the indicated antibodies. (B) HEK-293T cells were treated with 10 mM metformin treatment for 48 h. PP2A-C and CDC25B were detected using immunofluorescence. The nucleus was labeled by DAPI staining. Images were taken at 100× magnification. (C) BT549 and HS578T cell lysates were subjected to immunoprecipitation with control IgG or anti-CDC25B antibody. The immunoprecipitates were blotted with the indicated antibodies. (D) BT549 and HS578T cell lysates were subjected to immunoprecipitation with control IgG or anti-PP2A-C antibody. The immunoprecipitates were blotted with the indicated antibodies. (E) CDC25B knockout BT549 cells were transfected with EV, WT CDC25B (WT), CDC25B S353A, CDC25B S353E or CDC25B C488S constructs for 48 h. Cell lysates were subjected to immunoprecipitation with control IgG or anti-Flag antibody. The immunoprecipitates were blotted with the indicated antibodies. (F) BT549 cells were treated with 20 mM metformin for 48 h. Cell lysates were subjected to immunoprecipitation with control IgG or anti-CDC25B antibody. The immunoprecipitates were blotted with the indicated antibodies. (G) CDC25B knockout BT549 cells were transfected with EV, FL CDC25B, CDC25B N-terminal domain (1–373 aa, NTD) or CDC25B C-terminal domain (374–580 aa, CTD) for 48 h. Cell lysates were subjected to immunoprecipitation with control IgG or anti-Flag antibody. The immunoprecipitates were blotted with the indicated antibodies. Data information: all data presented are a representation of N = 3 independent experiments.