Fig. 7

MNAT1 regulates p53 down-stream molecules. A, HEK293T cells were transfected with pSIN or pSIN-MANT1, and then MNAT1, p53, p21, cleaved caspase3, caspase3, cleaved PARP, PARP, BAX, RAD51, and Bcl2 were detected with Western-blotting. pSIN served as a control. B, HCT116 p53^+/+ and HCT116 p53^−/− cells were transfected with pLVX-shscramble, pLVX-shMNAT1^#1 or pLVX-shMNAT1^#2, respectively. pLVX-shscramble served as a control. MANT1, p53, p21, BAX, PARP, and RAD51 were detected. C, LoVo cells were transfected with pLVX-shMNAT1^#1, and HEK293T cells were transfected with pSIN-MANT1. And then the transfected cells were treated with DOX. The treated cells were evaluated MNAT1, PARP, p53 expression. GAPDH served as a loading control. D, LoVo cells were infected with pLVX-shMNAT1^#1 or pLVX-shMNAT1^#2, and HEK293T cells were transfected with pSIN-MANT1, and then treated with 5-Fu. The treated cells were evaluated MNAT1, p53, and Fas expression. E, The apoptosis of the treated cells was analyzed using flow cytometry, and apoptosis rates were conducted. a, shscramble plus 5-FU; b, shMNAT1^#1 plus 5-FU; c, shMNAT1^#2 plus 5-FU; d, blank vector plus 5-FU; e, pSIN-MNAT1 plus 5-FU; f, apoptosis rates of the above treated cell. DOX, Doxorubicin; 5-Fu, 5-fluorouracil; *, p < 0.05