Fig. 8: Inhibiting autophagic degradation of p53 reverses the TRIM39 deficiency-suppressed CRC progression.
a Western blot assay of p53 levels in TRIM39 stably knocked down HCT116 cells. b HCT116 cells were transfected with empty vector or FLAG-TRIM39 for 48 h and treated with or without CQ (50 nM) for the last 4 h. The levels of p53 were detected by western blot. c HCT116 cells were transfected with empty vector or FLAG-TRIM39 for 48 h and treated with or without CQ (50 nM) for the last 4 h. Cells were fixed and immunofluorescently stained for p53. Representative confocal microscopic images are shown. Scale bar, 20 μm. d The fluorescence intensity of p53 in c was calculated. Data are shown as mean ± SD. Ten images were scored. One-way ANOVA. *P < 0.05; ***P < 0.001; ns no significance. e TRIM39 stably knocked down HCT116 cells were infected with empty vector, Rab7WT, Rab7T22N, and Rab7Q67L overexpression lentivirus. The levels of p53 were detected by western blot. f Western blot analysis of p53 levels of tumor tissues from different groups in Fig. 1j. g Western blot analysis of p53 levels of tumor tissues from different groups in Fig. 6g. h Quantification (arbitrary units) of p53 levels relative to GAPDH in tumor tissues from f. Data are shown as mean ± SD. i Quantification (arbitrary units) of p53 levels relative to GAPDH in tumor tissues from g. Data are shown as mean ± SD. j Western blot assay of p53 levels in TRIM39-depleted HCT116 cells with p53 stably knocked down. k The colony-formation assay of TRIM39-depleted HCT116 cells with p53 stably knocked down. Data are shown as mean ± SD. n = 3 samples per group. One-way ANOVA. ***P < 0.001. l The migration assay of TRIM39-depleted HCT116 cells with p53 stably knocked down. The average number of cells per field was calculated. Data are shown as mean ± SD. n = 3 samples per group, four fields per sample. One-way ANOVA. ***P < 0.001.
