Extended Data Fig. 3. Trapped PARP1 is SUMOylated and ubiquitinated.
a. Input controls for Fig. 2c, showing the efficacy of MLN-7243 to inhibit ubiquitylation. Data shown represent 3 biological replicas. b. Reciprocal denaturing IP over PARP1-FLAG showed accumulation of trapped PARP1 ubiquitination in HEK293s. HEK293 cells were transfected with PARP1-FLAG-expressing construct for 24 hours then treated with 100 nM talazoparib/0.01% MMS and/or 10 µM CB-5083. Cells were lysed, chromatin was digested and then incubated with anti-FLAG beads. 4% of sample was harvested for input pre-incubation. Data shown represent 2 biological replicas. c. As in (C), but the immunoprecipitated proteins were analysed with an anti-K48 Ub chains recognising antibody. This experiment has been performed once. d. High exposure blot of PARP1 SUMOylation from Fig. 2d, red arrows show SUMOylated PARP1 in MMS treated samples. Data shown represent 2 biological replicas.