Figure 3. Nucleolytic Incision of ICL Lesions by MutLα (MLH1-PMS2) (A) Schematic of nicking sites engineered into ICL pBS plasmids (see also Figure S3A). (B) Quantification of ICL repair in HSS of unnicked, 5′ or 3′ nicked plasmids at 90 min. Results represent mean ± SEM from n = 6 independent experiments. (C) Plasmid pull-down assay and WB of SJG-136-treated plasmids in mock- or MSH2-depleted extract. (D) MLH1-PMS2 immunodepletion. Mock- and MLH1-depleted HSS was analyzed by WB. (E) Quantification of ICL repair in mock-depleted, MLH1-depleted, or MLH1-depleted HSS supplemented with recombinant MutLαWT (n = 12) or MutLαn.d (n = 7) at 90 min. Results represent mean ± SEM independent experiments. (F) Quantification of ICL repair in HSS with overexpression of buffer, MutLαWT, or MutLαn.d. at 90 min. Results represent mean ± SEM from n = 5 independent experiments. See also Figure S3.
Image published in: Kato N et al. (2017)
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