Fig. 3: Polθ suppresses RAD52-mediated toxicity.
a RNF168 foci, b FLAG-RAD52 foci in asynchronous cells with control (−) or Polθ (+) siRNA. n = 120 cells from 3 biological replicates. c RAD52 and γH2AX foci in asynchronous Brca1C61G/C61G 53bp1−/− cells treated with control or Polθ siRNA. Scale bars represent 10 µm. d % γH2AX foci colocalised with RAD52 foci in asynchronous cells treated with control (−) or Polθ (+) siRNA. n = 80 cells from 2 biological replicates. e Colony survival of MEFs treated with Polθ, or control siRNA (−), and RAD52 siRNA. n = 4 biological replicates. f RAD51 foci in MEFs treated with IR (2 Gy) and Polθ siRNA, control (−) and RAD52 siRNA, fixed after 3 h. n = 150 cells from 3 biological replicates. g RAD52 western blot after 8 nM siRNA in Brca1C61G/C61G 53bp1−/− cells. Quantification relative RAD52 in control-treated cells, n = 2 biological replicates. h Colony survival of MEFs treated with NTC or Polθ siRNA and RAD52 inhibitor 6-OHD or vehicle (−). n = 3 biological replicates. i RAD51 foci in IR (2 Gy) -treated MEFs with RAD52 inhibitor 6-OHD or vehicle (−), fixed 3 h after exposure. n = 150 cells from 3 biological replicates. j Survival of CAL51 cells treated with Polθ, BRCA1 siRNA or both, and RAD52 inhibitor 6-OHD or vehicle (−). n = 3 biological replicates. k Quantification of FLAG-RAD52 foci in cells treated with Polθ (+), or control siRNA (−) and 0.15 μM 6-OHD or vehicle. n = 100 cells from 3 biological replicates. l Number of breaks or gaps per metaphase spread, from cells treated with control (−) or Polθ (+) siRNA and RAD52 inhibitor. n ≥ 80 metaphases from 3 biological repeats. Data are mean ± SEM. m Micronuclei after treatment with Polθ, or control siRNA (siNTC) with and without RAD52 inhibitor 6-OHD. n = 600 cells from 2 biological replicates. In all cases, Data shown are mean ± SEM. All statistical analysis in this figure was performed using a two-tailed Student’s t Test, without adjustment for multiple comparisons. Source data are provided as a Source Data file.
