Fig. 1: Overview of PALB2 VUSs examined in our study and validation of the U2OS/DR-GFP homologous recombination (HR) reporter system. | npj Breast Cancer

Fig. 1: Overview of PALB2 VUSs examined in our study and validation of the U2OS/DR-GFP homologous recombination (HR) reporter system.

From: Functional assessment of missense variants of uncertain significance in the cancer susceptibility gene PALB2

Fig. 1

a Schematic representation of the PALB2 protein and the position of the previously identified PALB2 VUSs. The p.L35P pathogenic variant was also included and highlighted in red. C.C., coiled-coil domain; ChAM, chromatin-association motif; WD40, WD40-repeats; NES, nuclear export sequence. b, c siRNA-mediated knockdown efficiency of endogenous PALB2 in U2OS/DR-GFP cells was evaluated by quantitative PCR and western blotting. Data represent the mean percentage (± SEM) of PALB2 mRNA relative to siCTL from three independent experiments. Statistical significance was analyzed by Student’s t-test. ***P < 0.001. d Schematic representation of the siRNA-resistant PALB2 cDNA containing eight silent base changes. e Representative plots for the HR activity of PALB2. The HR activity of U2OS/DR-GFP cells was first deprived after siRNA knockdown and then rescued with the siRNA-resistant PALB2-WT construct by transient transfection. GFP-positive cells were quantified by flow cytometry. EV Empty vector, WT Wild type.

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