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. 2020 Feb 10;11:819. doi: 10.1038/s41467-020-14563-y

Fig. 2. 53BP1 depletion rescues PALB2 focus formation in BRCA1-deficient cells.

Fig. 2

a Quantification of Venus-PALB2 IRIF in RPA focus-positive RPE1 cells. Two independently generated RPE1 Venus-PALB2 cell lines (#1 and #15) were siRNA-depleted for indicated proteins, exposed to 6 Gy of IR and 6 h later, fixed and stained with anti-GFP and anti-RPA2 antibodies. Imaging and IRIF quantifications were performed in three independent experiments, using OPERA Phoenix HT microscope. b Representative images, acquired on OPERA Phoenix HT microscope, of RPE1 cells with endogenously Venus-tagged PALB2 gene. The cells were stained with anti-GFP (to enhance the signal of the Venus tag) and anti-RPA2 antibodies. Scale bar, 50 µm. c Venus-PALB2 association with RPA filaments in cells depleted for 53BP1. RPE1 cells expressing endogenously tagged Venus-PALB2 were depleted for BRCA1 and/or 53BP1, irradiated with 6 Gy of IR and, 8 h later, processed for immunofluorescence analyses. Images were acquired using super-resolution 3D-SIM OMX microscope. Scale bar, 5 µm. Graphs to the right of the images represent distribution of relative frequencies of Venus-PALB2 foci numbers adjacent to each RPA focus. Source data are provided as a Source Data file.