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. 2019 Sep 5;116(39):19464–19473. doi: 10.1073/pnas.1903188116

Fig. 5.

Fig. 5.

PHF2 depletion leads to DNA damage and genome instability. (A) Immunostaining of NSCs expressing shControl (I), shPHF2 (II), shPHF2 together with PHF2 WT (III), and shPHF2 together with PHF2 (249H > A) (IV). Cells were fixed and stained with PHF2 and γH2Ax antibodies and DAPI. Enlarged images are showing individual cells. More than 30 cells were quantified. Data shown are representative of 2 to 3 biological independent experiments (Scale bar: 20 μm.) Boxplots represent the number of γH2Ax foci/cell. *P < 0.05; ***P < 0.001 (Student’s t test). (B) Immunostaining of a representative enlarged cell expressing shControl, shPHF2, and shPHF2 together with PHF2 (249H > A). Cells were fixed and stained with γH2Ax and S9.6 antibodies and DAPI. More than 30 cells were quantified. Data shown are representative of 2 to 3 biological independent experiments (Scale bar: 5 μm.) Boxplots represent the number of S9.6 foci/cell. ***P < 0.001 (Student’s t test). (C) Formation of multinucleated cells and segregation defects are shown for shControl and shPHF2 cells. A total of 50 to 100 cells were quantified. Data shown are representative of 4 independent experiments (Scale bar: 5 μm.) Boxplots represent the percentage of multinuclear cells. ***P < 0.001 (Student’s t test).