Fig. 2

ROS induced by mutant p53 proteins are critical to mediate their oncogenic proprieties. a, b, c AsPC1-p53 null cells were transfected with R175H or R273H vector, or mock control, and concomitantly treated with 7 mM NAC for 24 h. a Intracellular ROS level was evaluated analyzing DCF fluorescence intensity using a multimode plate reader. b Cell proliferation was measured by Crystal Violet assay and c apoptosis was determined by the annexin V/FITC binding assay. Statistical analysis: *p < 0.05; P values were calculated with two-tailed t test. R175H vs R175H + NAC and R273H vs R273H + NAC. d AsPC1-p53 null cells were transfected with R175H and R273H vector, or negative control, and concomitantly treated with 7 mM NAC and 1 µM GEM for 24 h. Cell proliferation was measured by Crystal Violet assay. Statistical analysis: *p < 0.05; P values were calculated with two-tailed t test. R175H + GEM vs R175H + NAC + GEM and R273H + GEM vs R273H + NAC + GEM. e AsPC1-p53 null cells were transfected with mock vector or with vector to express R175H mutant p53. After 24 h cells were treated with 100 µM H₂O₂ for further 24 h. Cell proliferation was measured by Crystal Violet assay. Statistical analysis: *p < 0.05 or ** p < 0.01 P value were calculated with two-tailed t test. R175H vs mock. f Kaplan–Meier curves of TTFT for CLL patient subgroups defined by mutational status of TP53 gene: wild-type (TP53-wt; n = 16) or mutated (TP53-mut; n = 10). g Comparison of reduced-thiol levels (detected by ThiolTracker probe) between CLL patients harboring TP53 gene wild-type (TP53-wt; n = 4) or mutated (TP53-mut; n = 3). Statistical analysis was performed using the Student’s t test. Data are expressed as mean ± SEM. *: P < 0.05