DSBs at the nuclear lamina are positionally stable. (A) Immunofluorescence of HT1080 cells expressing Dam-LaminB1 and m6A-Tracer 2 h after treatment (or not) with 50 ng/mL neocarzinostatin (NCS) for 15 min. (B) Box plot of GFP intensity ratios of the signal in the nucleoplasm versus the signal at the nuclear envelope in a HT1080-derived clonal cell line expressing a Dam-LaminB1 and the m6A-Tracer. The number of cells analyzed per condition was 20. For statistical analysis, χ2 tests were performed. (n.s.) Nonsignificant. (C) dSTORM microscopy images of LADs (green) and laminB (left panel; red) or TPR (right panel; red) in the absence (top panel) or presence (bottom panel) of DNA damage (100 ng/mL NCS for 15 min and released for 2 h) in HT1080 cells expressing Dam-LaminB1 and m6A-Tracer. Images were taken from the bottom of the cells to allow better resolution of nuclear pores. Corresponding colocalization and the ratio of positive over negative colocalization events are displayed at the right. The mean ratios for all nuclei analyzed (n ≥ 8) are displayed above.
