Fig. 6

MNAT1 shortens half-life time of p53. A, HEK293 cells were transfected with pSIN or pSIN-MNAT1, and then treated with 20 mg/ml CHX for the indicated time points. pSIN served as a control. MANT1 and p53 expressions were detected using Western-blotting (a). Quantitative analysis was conducted on p53 level in Western-blotting, the half-life time of p53 was calculated (b). B, LoVo cells were transfected with pLVX-shMNAT1^#1, p53 and MNAT1 expressions in the transfected cells were detected (a). Quantitative analysis was conducted on p53 level, and the half-life time was also calculated (b). C, HEK293 cells were transfected with pSIN-MNAT1, and LoVo cells were transfected with pLVX-shMNAT1^#1 or pLVX-shMNAT1^#2. p53 was immunoprecipitated, and MDM2 was detected in the immunocomplexes and WCL using Western-blotting (a). Quantitative analysis was conducted on p53 and MNAT1 expressions (b). D, MANT1-HEK293 cells were transfected with shMDM2, and p53, MDM2, MNAT1 expressions were detected in the transfected cells. CHX, cycloheximide; IP, immunoprecipitation; MGM2, murine double minute 2. WCL, whole cell lysate. *, p < 0.05