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. Author manuscript; available in PMC: 2016 Dec 17.
Published in final edited form as: Nature. 2015 Dec 9;528(7582):422–426. doi: 10.1038/nature16142

Figure 3. USP11 opposes the activity of CRL3-KEAP1.

Figure 3

a, Normal IgG or PALB2 immunoprecipitation (IP) of extracts derived from CPT-treated 293T cells of the indicated genotypes transfected with GFP-USP11 constructs. EV, empty vector; WT, wild type; CS, C318S. b, Clonogenic survival assays of 293T cells of the indicated genotypes treated with olaparib (mean ± s.d., N≥3). c, Normal IgG or PALB2 IP of extracts derived from CPT-treated 293T cells of the indicated genotypes. d, Immunoblots of deubiquitylation reactions containing ubiquitylated HA-tagged PALB2 (1-103) and increasing concentrations of GST-USP11 or its C318S (CS) mutant. USP2 was used as a control. e, Cell cycle-synchronized U2OS cells were irradiated (20 Gy dose) and processed for immunoblotting. f, Immunoblots of extracts from irradiated U2OS cells transfected with the indicated siRNAs. g, Fluorescence micrographs of G1-synchronized and irradiated (20 Gy) 53BP1Δ U2OS cells transfected with the indicated siRNAs. The percentage of cells with more than 5 γ-H2AX-colocalizing BRCA2 foci is indicated (mean ± s.d., N=3).