Fig. S2

The HR factors are specifically recruited at transcriptionally active damage sites in both U2OS TRE and U2OS 263 cell lines. (A) Recruitment of LIG4, BRCA1, CtIP, and BRCA2 at TA-KR and tetR-KR damage sites in U2OS TRE cells 10 min after light irradiation is shown. (B) U2OS TRE cells were cotransfected with GFP tagged RAD52 or RAD51C and tetR-cherry or TA-cherry; 24 h after transfection, cells were illuminated with fluorescent light for 10 min. There is no recruitment of these factors at sites of tetR-cherry or TA-cherry. (C) The recruitment of the HR factors in U2OS 2-6-3 cells. U2OS 263 cells were cotransfected with GFP tagged RAD52, or RAD51C and tetR-cherry/KR or TA-cherry/KR; 24 h after transfection, cells were illuminated with fluorescent light for 10 min. RAD51 was stained with antibody. The recruitment of these factors specifically at sites of TA-KR is shown. (D) Quantification of average fold increase of foci intensity (n = 10) of the NHEJ factor at sites of tetR-KR or TA-KR at the indicated recovery time after damage. Error bars indicate the SEM of three independent experiments.