Extended Data Fig. 4: Mutations of D and E residues in CycT1 IDR into A does not diminish MNNG-induced CycT1 PARylation; PARG treatment generates a homogenous pool of MARylated CycT1.

a, HeLa cells expressing the indicated CycT1-F proteins were untreated or treated with MNNG. The anti-Flag immunoprecipitates (IP) from whole cell extracts (WCE) were analyzed by Western blotting (WB). b, The immobilized af1521-Strep was incubated with WCE of HeLa cells untreated or treated with MNNG or/and PARG as indicated. The input and bound proteins were analyzed by WB. c, A schematic representation displaying PARG’s hydrolysis activity toward the PAR chain to leave behind a mono-ADP-ribose unit of 541.06 Dalton (Da). d, The Af1521-enriched CycT1-IDRL17A was untreated or treated with PARG in vitro and then analyzed by WB. All Western blots are representative of three independent experiments. Gel source data are available online.