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. 2023 Aug 7;120(33):e2302103120. doi: 10.1073/pnas.2302103120

Fig. 4.

Fig. 4.

Effect of the components of DNA mismatch repair on FAN1 activity. (A) (CAG)2 extrusion harboring circular DNA substrate was incubated with FAN1 or (FAN1 D960A), PCNA, and RFC in the presence of increasing concentration of MutSβ (as indicated). Indirect end labeling was performed with Fwd1947 probe. (B) Percentage of FAN1 cleavage as in A based on four independent experiments with error bars representing SD. *P < 0.05, ***P < 0.001, ****P < 0.0001, 1-way ANOVA with post hoc Dunnett’s test (comparison to FAN1 in the absence of MutSβ). (C) Percentage of FAN1 cleavage of (CAG)2 extrusion-containing DNA substrate in the presence of PCNA and RFC and increasing concentration of MutLα (as indicated) and analyzed as in A. Quantification based on five independent experiments with error bars represent SD. ns- not significant, one-way ANOVA with post hoc Dunnett’s test (comparison to FAN1 in the absence of MutLα). (D) (CAG)2 extrusion DNA substrate was incubated with FAN1 or (FAN1 D960A), PCNA, RFC, MutSβ, and MutLα as indicated. Products of the reaction were digested with PspFI and resolved on 10% polyacrylamide gel containing 8 M urea. Indirect end labeling was performed with Fwd3028 probe. The size marker was generated by digestion of mr77 4xLacO with AlwNI, XbaI, or AatII. The distance of each DNA fragment from the nick is indicated on the side. The experiment was repeated twice with similar outcome. See also SI Appendix, Fig. S4.