1. Expression of SYCE2 activates the DSB repair pathway.
Expression of SYCE2 activates the DSB repair pathway. (A) Immunofluorescence visualization of γH2AX foci (red) in MCF7 cells transfected with nontargeting control siRNA (upper panel) and those transfected with SYCE2-targeting siRNA (lower panel). Scale bar, 20 μm. (B) Percentages of cells containing more than five large γH2AX foci in MCF7 cells transfected with nontargeting control siRNA and those transfected with SYCE2-targeting siRNA, in unirradiated states and at 1, 8, and 24 h after 1-Gy x-ray irradition. A total of 100 cells were examined for each cell clone and at each time point. Columns and bars represent the mean of three independent experiments and SD, respectively. (C) Immunofluorescence visualization of 53BP1 foci (green) in MCF7 cells transfected with nontargeting control siRNA (upper panel) and those transfected with siRNA for SYCE2 (lower panel), treated at 2 h after 0.2-Gy x-ray irradiation. Scale bar, 20 μm. (D, E) Percentages of cells containing three or more large 53BP1 foci in unirradiated states and at 2 h after 0.2-Gy x-ray irradiation. Columns and bars represent the mean of three independent experiments and SD, respectively. A total of 100 cells were examined for each cell line. (F) NHEJ activity measured by in vivo DNA-ligation assay. The relative percentages of recirculation in MCF7 cells transfected with SYCE2-targeting siRNA compared with those in control cells were evaluated in three independent experiments. Columns and bars represent the mean and SD. (G) Immunofluorescence visualization of RAD51 foci (green) in MCF7 cells transfected with nontargeting control siRNA (upper panel) and in MCF7 cells transfected with siRNA for SYCE2 (lower panel), stained at 2 h after 8-Gy x-ray irradiation. Scale bar, 20 μm. (H, I) Percentages of cells containing more than five large RAD51 foci. Columns and bars represent the mean of three independent experiments and SD, respectively. A total of 100 cells were examined for each cell line. (J) HR repair activity measured by the DR-GFP assay. The relative percentages of the GFP-positive cells in HeLa-DRGFP cells transfected with SYCE2-targeting siRNA compared to those transfected with nontargeting control siRNA were evaluated in three independent experiments. Columns and bars represent the mean and SD, respectively.
Noriko Hosoya et al. LSA 2018;1:e
© 2018 Hosoya et al.