Fig. 7

Functional studies of RAN and XRCC6 inactivation. a Western blot analysis of nuclear/cytoplasmic fraction in PD(+)RAN-1 and PD(+)RAN-2 before and after RAN silencing. The nuclear translocation of pEGFR and its transcriptional targets (Cox2 and Cyclin D1) was examined. GAPDH and Lamin A/C were used as a cytoplasmic and nuclear specific loading control, respectively. b Confirmation of RAN suppression by two sgRNAs (sgRAN-1 and sgRAN-2) in two cells (PD(+)RAN-1 and PD(+)RAN-2). a, b Images of the full uncropped scans are provided in Additional file 1: Figure S10. c Immuno-fluorescence images for Rad51 (green) in irradiated PD(+)XRCC6 (upper) and PD(−)XRCC6 (lower) cells before and after XRCC6 inactivation (control vs sgXRCC6). DAPI staining results (blue) are shown side by side. d A graph showing the ratio (sgXRCC6/control) of the number of Rad51 foci per cells. The mean and standard error were obtained from 5 fluorescence images. e Scatter plots showing the rate of HR repair based on the DR-GFP/I-SceI assay in irradiated PD(+)XRCC6 and PD(−)XRCC6 cells. GFP-positive cells in the marked zones indicate a population that underwent HR-mediated DSB repair of GFP reporter plasmids. f A graph showing the ratio (sgXRCC6/control) of the GFP-positive cells. The mean and standard error were obtained from 5 experiments after excluding the maximum and minimum