Figure 6.

Mthfd2 Modulates HR Repair by Regulating EXO1 Phosphorylation

(A and B) IF staining for γ-H2AX in Mthfd2 KD mESCs at indicated times post CPT treatment. Representative images (A) and quantification of the average number of γ-H2AX foci per cell (B) (n = 50 nuclei) are shown. DAPI was used to indicate the nuclei. Scale bar, 50 μm.

(C and D) IF staining for γ-H2AX in Exo1 KD mESCs at indicated times post CPT treatment. Representative images (C) and quantification of the average number of γ-H2AX foci per cell (D) (n = 50 nuclei) are shown. DAPI was used to indicate the nuclei. Scale bar, 50 μm.

(E and F) Immunofluorescence staining for γ-H2AX in RO-3306 (RO)-treated mESCs at indicated times post CPT treatment. Representative images (E) and quantification of the average number of γ-H2AX foci per cell (F) (n = 50 nuclei) are shown. DAPI was used to indicate the nuclei. Scale bar, 50 μm.

(G) DNA damage levels in OE Cdk1-Mthfd2 KD mESCs were evaluated by the comet assay at indicated times post CPT treatment. Representative images (upper panels) and quantification of the mean olive tail moment (lower panel) (n = 50 nuclei) are shown. Scale bar, 100 μm.

(H and I) IF staining for RAD51 in Mthfd2 KD mESCs at indicated times post CPT treatment. Representative images (H) and quantification of the average number of Rad51 foci per cell (I) (n = 50 nuclei) are shown. DAPI was used to indicate the nuclei. Scale bar, 20 μm.

All data are pooled from three independent experiments (mean ± SEM). ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001 (Student's t test) compared with the control. See also Figure S6.