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. 2023 Jul 9;19(11):3544–3557. doi: 10.7150/ijbs.82566

Figure 3.

Figure 3

The synthetic lethality of PTEN and MDM2 is mediated by p53. A. Immunoblots showing AKT and MDM2 phosphorylation levels in PTEN-isogenic HCT116 cells. B. Immunofluorescence analysis of MDM2 cellular distribution, and the nuclei were labeled by DAPI. C, D. p53 protein expression levels in HCT116 and RKO PTEN-isogenic cells. Immunoblot showing MDM2, AKT, p-AKT (ser473) and p53 protein expressions with DMSO, 5 μM and 10 μM CGM097 treatment in HCT116 PTEN-isogenic cells (E) and RKO PTEN-isogenic cells (F). Quantification results of p53 protein expressions in HCT116 (G) and RKO (H) PTEN-isogenic cells, which were relative to the loading control (GAPDH) and normalized to the corresponding control group. **P < 0.01 between two indicated groups (t test). Rescue experiments with p53 inhibitor pifithrin-α in HCT116 PTEN-/- cells (I) and RKO PTEN-/- cells (J). The cells were cotreated with pifithrin-α and CGM097 for 72 h, and the cell viability was measured by AlamarBlue assay. K-L. HCT116 and RKO PTEN-/- cells were transfected with p53 siRNA and co-incubated with CGM097 for 72 h. The cell viability was measured as the same method above. M. Immunoblots indicating p53 siRNA transfection efficiency.