Fig 2: Cohesin- and PBAF-Dependent Transcriptional Repression at DNA Double-Strand Breaks Occurs in Both G1 and G2 Phases(A and C) Representative images of cells expressing GFP-BAF180 (A), GFP-SA2 (C), or Cdt1-RFP (to identify G1 phase cells) as indicated following laser microirradiation.(B and D) Quantification of GFP-BAF180 (B) or GFP-SA2 (D) recruitment to laser-microirradiation-induced damage in G1 cells (Cdt1-RFP positive) or cells outside of G1 (Cdt1-RFP negative). Data represent the relative mean signal intensity ± SEM for n = 6 (B) or n = 7 (D) biological repeats. At least 42 cells were analyzed in total for each construct per cell-cycle phase.(E) Quantification of transcription in CENPF-positive (G2 phase) U2OS reporter cells treated with the indicated siRNA (NTC, non-targeting control) with or without induction of the FokI endonuclease. Data are presented as the mean ± SD. More than 40 cells were analyzed per condition per repeat (n = 3 biological repeats).(F) Representative images of U2OS reporter cells analyzed in (E).(G) Quantification of transcription in cyclin-D1-positive (G1 phase) U2OS reporter cells treated with the indicated siRNAs. Data are presented as the mean ± SD. More than 40 cells were analyzed per condition per repeat (n = 3 biological repeats).(H) Representative images of U2OS reporter cells analyzed in (G).(I) Quantification of ?H2AX foci clearance following exposure to 1.5 Gy IR in G1 phase (cyclin D1 positive) U2OS cells treated with the indicated siRNA. Data are presented as mean ± SD; n = 3 biological repeats.*p < 0.05, **p < 0.01 using paired Student’s t test. See also Figure S2.