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. 2022 Aug 21;12(14):6160–6178. doi: 10.7150/thno.70814

Figure 3.

Figure 3

RYR2 interacted with MDM2. (A) Immunofluorescence analysis of the localization of RYR2/MDM2 in YES2 and KYSE150 cells. (B) Co-immunoprecipitation of RYR2 and MDM2 by RYR2 antibody. The cellular lysate was used to monitor the expression of MDM2 and RYR2 by western blotting (input). From the remaining cellular lysate, RYR2 was immunoprecipitated and associated MDM2 was detected by western blotting. Precipitation with IgG was performed as a negative control. (C) Ubiquitination of endogenous p53 was analyzed by immunoprecipitation with p53 antibody and followed by western blot analysis. YES2 and KYSE150 cells were knockdown of RYR2 and after 42h the cells were treated with MG132 (10 µM) for another 6h, then the cells were lysis and used for IP assay. (D) Co-immunoprecipitation of p53 and MDM2 by p53 antibody. YES2 (left) /KYSE150 (right) cells were lysed and used to monitor the expression of MDM2 and p53 by western blotting (input). From the remaining cellular lysate, p53 was immunoprecipitated and associated MDM2 was detected by western blotting. (E) Western blot analysis of the expression level of p53, p21 and GAPDH in YES2 and KYSE150 cells transfected with siRYR2 or siCtrl for 48h.