PMC

(A) MCF-7 cells were treated with NC, 80 MOI Ad-p66Shc and 160 MOI Ad-p66Shc for 48 h. DCFH-DA staining showed that p66Shc significantly induced intracellular ROS. *p < 0.05, ***p < 0.001 vs Negative control. (B) MCF-7 cells were infected with 100 MOI Ad-p66Shc or negative control for 48 h. The upregulation of p66Shc also increased the level of 8-OHdG. The data represent the means ± SEM, n = 3 independent experiments. **p < 0.01 vs NC (NC: negative control).

(A) MCF-7 cells were infected with various concentrations of Ad-p66Shc or negative control (20, 40, 80, 160, or 320 MOI) for 48 h. (B) MCF-7 cells were infected with 100 MOI Ad-p66Shc for different amounts of time (12, 24, 36, 48, or 60 h), and cell viability was detected using the MTT method. The data represent the means ± SEM, n=6 independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs NC (NC: negative control).

MCF-7 cells were infected with Ad-p66Shc or negative control for 48 h. (A) The cell extracts were prepared and analyzed by Western blot with the corresponding antibodies. The quantification of the digital images was performed using ImageJ software. (B) MCF-7 cells were stained with annexin V-FITC and PI and analyzed by flow cytometry. The data represent the means ± SEM, n = 3 independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs NC (NC: negative control).

(A) Schematic presentation of p66Shc homologous recombination with pAdenoX-CMV. CH1: collagen-homology region; PTB: phosphotyrosine-binding domain; SH2: Src-homology2 domain; P_{CMV IE}: cytomegalovirus immediate early promoter; SV40 polyA: simian virus 40 polyA signals; ITR: inverted terminal repeat; AMP: ampicillin. (B) HEK293A, HUVECs, HeLa and MCF-7 cells were infected with Ad-p66Shc or negative control (NC) for 48 h. The expression of p66Shc protein was detected by Western blot.

MCF-7 cells were infected with Ad-p66Shc or negative control for 48 h. (A) After treatment, the cells were examined by flow cytometer for cell cycle analysis. (B) The cell extracts were prepared and analyzed by Western blot with the corresponding antibodies. The quantification of the digital images was performed using ImageJ software. The data represent the means ± SEM, n = 3 independent experiments. **p < 0.01, ***p < 0.001 vs NC (NC: negative control).