Fig. 3: PARP1 expression abolishes DNA damage induced by PLA2R1.
From: PLA2R1 promotes DNA damage and inhibits spontaneous tumor formation during aging
A–D MRC5 normal human cells were infected with a control, PLA2R1-, or PARP1-encoding retroviral vectors as indicated and selected. A RNA was prepared and RTqPCR performed against PLA2R1 or PARP1 and normalized against GAPDH levels. The results shown are mean ± SEM of three independent experiments. P value was calculated using Student’s t-test. *p < 0.05; **p < 0.01; ***p < 0.001. B, C Cells were fixed in methanol and immunofluorescence staining against 53BP1 (B) and γH2AX (C) was performed with indicated antibodies. Images were taken using a Nikon Fluorescence microscope and quantified with the Focinator software. More than 50 nuclei were analyzed per condition. The p value was calculated using the Mann–Whitney test. *p < 0.05; **p < 0.01; ***p < 0.001. D Comet assay for assessing DNA breaks was performed. After migration, the slides were stained with SYBR Safe. Tail moments (1/4 tail length DNA in the tail/total DNA) were analyzed using the Casplab software. A least 50 nuclei were analyzed per condition. The p value was calculated using the Mann–Whitney test, ***p < 0.001.
