Fig. 2: SIRT5 suppresses p53 target gene expression in HCT116 cells upon DNA damage.
From: Repression of p53 function by SIRT5-mediated desuccinylation at Lysine 120 in response to DNA damage
A, B Overexpression of SIRT5 in HCT116 cells caused a reduction of p21 (A) and MDM2 (B) mRNA upon Dox (Doxorubicin) treatment (1 μM, 24 h), as revealed by qPCR analysis. DMSO was used as control. Data show mean ± SEM; Student’s two-tailed t-test; Data from three independent experiments. C Overexpression of SIRT5 in HCT116 cells caused a reduction of the protein levels of p21 and TIGAR upon Dox treatment (1 μM, 24 h), as revealed by Western blot analysis. DMSO was used as control. D–G qPCR analysis of p21 (D), PUMA (E), TIGAR (F), and MDM2 (G) mRNA in SIRT5-deficient or SIRT5-intact HCT116 cells (SIRT5−/− or SIRT5+/+) treated with or without Dox (1 μM, 24 h). DMSO was used as control. Data show mean ± SEM; Student’s two-tailed t-test; Data from three independent experiments. H Western blot analysis of p21 and TIGAR protein level in SIRT5-deficient or SIRT5-intact HCT116 cells (SIRT5−/− or SIRT5+/+) treated with or without Dox (1 μM, 24 h). DMSO was used as control. I, J qPCR analysis of p21 (I), and MDM2 (J) mRNA in SIRT5-deficient or SIRT5-intact HCT116 cells (SIRT5−/− or SIRT5+/+) treated with or without Nutlin-3 (5 μM, 24 h). DMSO was used as control. Data show mean ± SEM; Student’s two-tailed t-test; Data from three independent experiments. K Western blot analysis of p21 and TIGAR protein level in SIRT5-deficient or SIRT5-intact HCT116 cells (SIRT5−/− or SIRT5+/+) treated with or without Nutlin-3 (5 μM, 24 h). DMSO was used as control. L, M qPCR analysis of p21 (L), and MDM2 (M) mRNA in SIRT5-deficient or SIRT5-intact HCT116 cells (SIRT5−/− or SIRT5+/+) treated with or without 5-Fu (5-fluorouracil) (5 μg/ml, 24 h). DMSO was used as control. Data show mean ± SEM; Student’s two-tailed t-test; Data from three independent experiments. N Western blot analysis of p21 and TIGAR protein level in SIRT5-deficient or SIRT5-intact HCT116 cells (SIRT5−/− or SIRT5+/+) treated with or without 5-Fu (5 μg/ml, 24 h). DMSO was used as control.
