Fig. 5

NR2F2 mitigated cell senescence by reducing DNA damage. (A, C) The comet assay (A) and 8-OH-dG content analysis (C) were used to assess the extent of DNA damage in A549, MLE-12, and BEAS2B cells after transfection with control or NR2F2 overexpression plasmid for 48 h, followed by an additional 72 h of treatment with 0.02 u/ml bleomycin. (B, D) The comet assay (B) and 8-OH-dG content analysis (D) were employed to evaluate the extent of DNA damage in A549, MLE-12, and BEAS2B cells stably expressing control and NR2F2 knockdown plasmids. (E) The protein expression level of γH2AX in A549, MLE-12, and BEAS2B cells treated with 0.02 U/ml bleomycin for an additional 72 h after transfection with control or NR2F2 overexpression plasmid for 48 h were quantified using WB. (F) The protein expression level of γH2AX in A549, MLE-12, and BEAS2B cells stably expressing control and NR2F2 knockdown plasmids were measured using WB. (G) Representative immunofluorescence staining of γH2AX expression in A549 cells treated with 0.02 U/ml bleomycin for an additional 72 h after transfection with control or NR2F2 overexpression plasmid for 48 h. (H) Representative immunofluorescence staining of γH2AX expression in A549 cells stably expressing control and NR2F2 knockdown plasmids. Data are shown as the mean ± SD. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001