Fig. 7
From: TSPYL2 is a novel regulator of SIRT1 and p300 activity in response to DNA damage
TSPYL2 promotes DNA damage-induced apoptosis in a SIRT1- and p300- dependent manner. a U2OS cells were transfected with control or TSPYL2-specific siRNAs and the induction of pro-apoptotic markers (cleaved PARP and cleaved caspase-3) was evaluated 30 h after etoposide treatment (20 µM) by western blot. b Control and TSPYL2-depleted U2OS cells were treated or not with etoposide for 30 h and scored for viability with trypan blue staining. Values are mean ± s.d. of three independent experiments. c U2OS cells were transfected with control, TSPYL2 and SIRT1-specific siRNAs and the induction of PARP cleavage was evaluated 30 h after etoposide treatment (20 µM) by western blot. The fold induction of cleaved PARP relative to loading control (vinculin) is indicated. d U2OS cells were transfected and treated as in c and the percentage of dead cells was determined by trypan blue exclusion test. Values are mean ± s.d. from three independent experiments. e U2OS cells transfected with MOCK or p300-encoding vectors were silenced for TSPYL2 and the levels of cleaved PARP were determined 30 h after etoposide treatment (20 µM) by western blot. The fold induction of cleaved PARP relative to vinculin-loading control is reported. f U2OS cells were transfected and treated as in e and the percentage of dead cells was determined by trypan blue exclusion test. Values are mean ± s.d. from three independent experiments. g colonies formation assays were performed on U2OS cells transfected with control or TSPYL2 siRNAs and treated with increasing doses of etoposide. Colonies were counted and indicated as percentages relative to the untreated samples. * p = 0.01; **p = 0.008
