Figure 1
Isolation of cytoplasmic and nuclear p53 protein for A549 cells after treated with interleukin 7 (IL-7), anti-IL-7 receptor antibody (A-IL-7R) for western blot analysis. Then total RNA was isolated for real-time quantitative PCR (RT-qPCR) assay after incubated with IL-7 for different time. (a) A549 cells were treated with IL-7 at 24 ng/ml for 8 h and 12 h, then p53 nuclear and cytoplasmic protein were detected. Lamin B was used as nuclear loading control and GAPDH was used for cytoplasmic loading control. Cytoplasmic Lamin B and nuclear GAPDH were detected to validate nuclear protein was not contaminated by cytoplasmic protein and cytoplasmic protein were not contaminated by nuclear protein. Bar show the cytoplasmic and nuclear p53 western blot analysis. (b) A549 cells were treated with IL-7 and A-IL-7R plus IL-7 for 12 h, then nuclear and cytoplasmic p53 protein were detected by western blot. Cytosolic Lamin B and nucleus GAPDH was detected as negative control. Bar show the cytoplasmic and nuclear p53 western blot analysis. (c) After incubated with IL-7 at different time (0 h, 8 h, 12 h), p53 mRNA level was detected by RT-qPCR, all the bar graph expressed as Mean ± SD, n = 3. *represent for P < 0.05 versus negative control (NC). **represent for P < 0.01 versus NC.
