Fig. 6 | Cell Death & Differentiation

Fig. 6

From: βarrestin-1 regulates DNA repair by acting as an E3-ubiquitin ligase adaptor for 53BP1

Fig. 6

βarr1 forms a complex with Rad18/Rad6 promoting polyubiquitylation of 53BP1. a Immunoblot showing complex formation of βarr1, Rad18/Rad6, and 53BP1 in vitro. Purified 53BP1-myc was incubated with either GST or GST-βarr1 in the presence or absence of Rad18/Rad6. Proteins residing in the precipitates were subjected to western blot analysis and probed with the indicated antibodies. Presented data was cropped from the same blot image exposed for an equal time-period. b HEK293 cells were transfected with either control (Ctrl 2) siRNA or siRNA directed against Rad18 and 53BP1 levels were determined by immunoblotting. c HEK293 cells were transfected with either control (Ctrl) siRNA or siRNA directed against Rad18 and treated with 2 µg/mL bleomycin (BLM). Cell lysates were immunoprecipitated with anti-53BP1 antibody and levels of polyubiquitylated 53BP1 were assessed by western blot using an anti-ubiquitin (P4D1) antibody. All bars represent mean ± s.e.m of three independent experiments. d Lysates derived from WT and βarr1−/− MEFs treated with 2 µg/mL BLM were immunoprecipitated using an anti-53BP1 antibody, immunoblotted, and probed with an anti-ubiquitin (P4D1) antibody. All bars represent mean ± s.e.m. of three independent experiments

Back to article page