Fig. 3.

FAN1 directly interacts with ub-PCNA through its UBZ domain in vitro and in vivo. a GST, full-length GST-FAN1 WT and the UBZ* mutant were incubated with purified ub-PCNA and subsequently retrieved with glutathione beads. ub-PCNA in input and pull-downs was analysed by immunoblotting. b Chromatin-enriched fractions derived from U2OS cells inducibly-expressing eGFP-tagged FAN1-WT and the UBZ* mutant, treated or mock-treated with aTMS (2 μΜ; 24 h), were incubated with anti-eGFP affinity resin. Inputs and immunoprecipitates were analysed with the indicated antibodies. c Chromatin-enriched fractions derived from cells as in b, treated or mock-treated with aTMS (2 μΜ; 24 h) and transfected or not with siRNAs against RAD18, were incubated with an anti-PCNA antibody. The immunoprecipitates were analysed with antibodies against PCNA and FAN1