Fig. 7
From: Functional analysis of genetic variants in the high-risk breast cancer susceptibility gene PALB2
Functional analysis of damaging PALB2 variants in human cells. a CRISPR-LMNA HDR assay in siRNA-treated U2OS PALB2 knockdown cells expressing siRNA-resistant human PALB2 cDNA with the indicated variants (or an empty vector control, Ev). Data represent the mean percentage (±SD) of mRuby2-positive cells among the YFP-positive cells from three independent experiments (n > 300 YFP-positive cells per condition) relative to wild type (WT), which was set to 100%. b PARP inhibitor (PARPi) sensitivity assay using siRNA-treated HeLa PALB2 knockdown cells expressing siRNA-resistant human PALB2 cDNA with the indicated variants (or an empty vector control, Ev). Survival curves were determined after 72 h of PARPi treatment. Data represent the mean percentage of viability relative to untreated cells ( ± SD), which was set to 100%, of three independent experiments, each performed in triplicate. c Representative images of RAD51 foci 4 h after 2 Gy of ionizing radiation in siRNA-treated HeLa PALB2 knockdown cells expressing siRNA-resistant human PALB2 cDNA with the indicated variants (or an empty vector control, Ev). Scale bar: 5 µm. d Quantification of the results from (c). Scatter dot plot shows the number of RAD51 foci in cyclin A-positive cells expressing the indicated variant, with the horizontal lines designating the mean values (±SD) of three independent experiments (n > 200 cells per condition). e Quantification of the results from (c). Scatter dot plot shows the intensity of RAD51 foci in cyclin A-positive cells expressing the indicated variant, with the horizontal lines designating the mean values (±SD) of three independent experiments (n > 500 cells per condition). Source data are provided as a Source Data file.
