Extended Data Fig. 5: PARylation of PDHE1α by PARP1 is essential for its recruitment to DNA damage sites.
From: PARylated PDHE1α generates acetyl-CoA for local chromatin acetylation and DNA damage repair
a-b, Western blot showing PDHE1α PARylation and the effect of individal potential PARylation site mutation (a) and combined mutations (b) on chromatin accumulation. PDHE1α-deficient HeLa cells were transfected with PDHE1α WT and a mutant construct, irradiated with 3 Gy IR and different fractions were harvested immediatedly. The cell lysates were subjected to immunoprecipitation using anti-HA agarose and analysed by western blotting. c, Dynamics of PDHE1α accumulation at DNA damage sites monitored by laser micro-irradiation-coupled live-cell imaging of HeLa cells expressing NLS-fused PDHE1α-WT or PDHE1α-6A constructs. GFP signal accumulation intensity at DSB stripes was quantified from three independent replicates (n = 20 for each group), the curve shows the mean ± s.e.m. Scale bars, 5 μm.
