Supplementary Figure 2: p73 enhances G6PD expression.
From: TAp73 enhances the pentose phosphate pathway and supports cell proliferation
(a) The pentose phosphate pathway and its link with glycolysis. FBP: fructose 1,6-biphosphate. PEP: phosphoenolpyruvate. TCA cycle: tricarboxylic acid cycle. (b) Expression of G6PD and TAp73 proteins in U2OS cells transfected with control siRNA or p73 siRNA, plus control vector or siRNA-resistant plasmid encoding Flag-TAp73. Western blots represent two independent experiments. (c) Semiquantitative RT-PCR analysis of ΔNp73 expression in HeLa, U2OS, H1299, and p53+/+ HCT116 cells. (d) Comparison of effect of p73 and ΔNp73 on G6PD expression in p73, ΔNp73 or control siRNA in HeLa cells by RT-PCR. (e) U2OS cells were infected with lentiviruses expressing a TAp73 shRNA or a control shRNA and cultured for the indicated time. The expression of G6PD, TAp73, and actin was analyzed by Western blot. The data represents three independent experiments. (f) mRNA expression in H1299 cells stably expressing control shRNA, p63 shRNA, or TAp73 shRNA was assayed by RT-PCR. (g) Quantitative RT-PCR analysis of G6PD expression in H1299 cells transfected with increasing amounts of control plasmid or plasmid expressing the indicated p53 family proteins. Data are means ± SD (n = 3 independent experiments). (h) A549 cells were treated with 1 μg/ml doxorubicin for 24 hour and protein expression was analyzed by Western blot. (i) Luciferase reporter assay of 293T cells transfected with p73 RE-luciferase construct or control construct, plus TAp73 or ΔNp73. Data are means ± SD (n = 3 independent experiments). (j) H1299 cells were transfected with increasing amounts of Flag-p53R175H, Flag-p53R273H, or Flag-TAp73α as indicated. Protein expression was examined by Western blot. The data represents two independent experiments.
