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. 2018 Feb 13;46(7):3468–3486. doi: 10.1093/nar/gky098

Figure 6.

Figure 6.

Involvement of 53BP1 and Rif1 in DNA damage repair by Pold3. (A) Immunofluorescence and quantification of Pold3 (Flag) and 53BP1 foci after treatment with etoposide for 2 h. 20 nuclei were randomly counted. (B) Co-localization of Rif1 and 53BP1 by immunofluorescence after exposure to 2.5 μM etoposide for 2 h. Right panel bar figure, 20 cells were counted. (C) Following exposure to 2.5 μM etoposide for 2 h, Pold3 KD (shPold3) ESCs (N33) exhibit more Rif1 foci by 12 h compared with shControl. Scale bar, 5 μm. n = 50 cells counted. (D) Increased sensitivity to ATR inhibitor in Pold3 KD (shPold3) ESCs than in shControl ESCs. ESCs were plated in 12-plates, and various concentrations of VE-821 were added in the culture medium, and number of cells counted at day 3. Alternatively, ESCs were plated in 12-plates, and 3 μM VE-821 was added in the medium. Cells were counted and replated into new plates every two days. Data were analyzed using Two-way ANOWA. Data represent mean ± SEM. ***P < 0.001.