Fig. 5
From: TSPYL2 is a novel regulator of SIRT1 and p300 activity in response to DNA damage
In response to DNA damage, SIRT1 inhibits p300 activity by deacetylation. a U2OS cells were transfected with siLUC or siTSPYL2 siRNAs and 48 h later p300 was immunoprecipitated before and after etoposide treatment. p300 acetylation was determined by western blot with a general anti-acetyl-lysine antibody. The mean fold induction ± s.d. of acetylated p300 relative to total p300 protein from three independent experiments is shown in the chart. IP, immunoprecipitates; PC, negative control. Input, total lysates. b p300 was immunoprecipitated from untreated and etoposide treated U2OS-SIRT1-WT or U2OS-SIRT1-KO cells. Levels of p300 acetylation were analyzed by western blot with a general anti-acetyl-lysine antibody. The mean fold induction ± SD of acetylated p300 normalized to immunoprecipitated p300 from three independent experiments is indicated in the graph. c U2OS cells were transfected with MOCK or SIRT1 encoding vectors and 48 h later p300 was immunoprecipitated before and after etoposide treatment. p300 acetylation was determined by western blot with a general anti-acetyl-lysine antibody. As above, the mean fold induction ± SD of acetylated p300 relative to immunoprecipitated p300 from three independent experiments is reported in the chart. PC, negative control. d U2OS cells were transfected with control or TSPYL2 siRNAs, treated with MG132 and then with etoposide. p53 was immunoprecipitated and its interaction with SIRT1 was analyzed by western blot. The fold induction of co-immunoprecipitated SIRT1 relative to immunoprecipitated p53 is indicated. IP, immunoprecipitates; PC, negative control; Input, total lysate
