Fig. 6: Mutational analysis of PARP1 serine auto-modification.
From: The regulatory landscape of the human HPF1- and ARH3-dependent ADP-ribosylome
a PARP1 auto-modification analysis, showing absolute modification abundance. n = 4 cell culture replicates, data are presented as mean values ± SEM. b As a, but showing relative modification abundance. c Immunoblot analysis highlighting the ADPr equilibrium in PARP1 wildtype (WT) and 3SA mutant cells, ±Olaparib pre-treatment at 10 μM for 1 h (“PARPi”), ± H2O2 treatment at 5 mM for 30 min. The arrow indicates histone ADPr. This experiment was performed as two independent biological replicates, with similar results. d Representative live-cell microscopy images, visualizing GFP-PARP1 recruitment following laser microirradiation. The scale bar represents 5 μm. e Quantification of recruitment kinetics of GFP-PARP1 to damage stripes, as monitored via full visual dissipation of accumulated PARP1 signal. Stratification classes and all stratified data are available in Supplementary Fig. 5. PARP1 kinetics were significantly different between WT and 3SA mutant in the absence of PARPi, at p = 1.0 × 10−5 for T = 1, p = 1.0 × 10−5 for T = 4, p = 7.9 × 10−5 for T = 7, p = 1.0 × 10−5 for T = 10, p = 2.4 × 10−4 for T = 13, and p = 0.017 for T = 16, as determined via chi-squared testing on all stratified data (Supplementary Fig. 5B). n = 109 cells per condition. Source data are provided as a Source Data file.
