Fig. 4: The ADP-ribosylome in HPF1 and ARH3 knockout cells.
From: The regulatory landscape of the human HPF1- and ARH3-dependent ADP-ribosylome
a Overview of the experimental design. U2OS cells, either wild-type (control), HPF1 knockout (KO), or ARH3 KO, were cultured in quadruplicate (n = 4), and either mock- or H2O2-treated at 1 mM for 10 min. ADPr sites were enriched using the Af1521 methodology, fractionated, and analyzed using high-resolution MS. n = 4 cell culture replicates. b Immunoblot analysis validating the knockout of HPF1 and ARH3. This experiment was performed as two independent biological replicates, with similar results. The same set of samples was loaded on multiple membranes as technical replicates, to allow readout using different antibodies. Ponceau-S loading controls for each membrane are available in the Source Data. c As b, but highlighting the ADPr equilibrium in control, HPF1 KO, and ARH3 KO cells, ± H2O2 treatment at 1 mM for 10 min. The arrow indicates histone ADPr. d Overview of the number of identified and localized ADPr sites. n = 4 cell culture replicates, data are presented as mean values ± SD. e As d, showing ADPr abundance. f Hierarchical clustering analysis of z-scored ADPr site abundances, visualizing the relative presence of ADPr sites across the experimental conditions. g Principle component analysis indicating the highest degree of variance between sample conditions. h Scaled Venn diagram depicting the overlap between ADPr sites in untreated cells. i As h, but for H2O2-treated cells. Source data are provided as a Source Data file.
