Figure 1

BCG modulates TNF-α-induced p53. (A) The expression of p53 in A549 cells treated with various concentration of TNF-α as indicated was analyzed using immunoblotting. Adriamycin (500 ng/ml) was used as positive regulator of p53 expression. (B) The p53 promoter luciferase construct was transfected into A549 cells and promoter activity was assayed at various concentrations of TNF-α as indicated. (C) A549 cells were infected with BCG for 12 h prior to treatment with TNF-α. p53 expression levels were assessed by immunoblotting. (D) A549 cells were transfected with p53 luciferase construct and change in p53 promoter activity with respect to indicated treatments was measured by luciferase assay. (E) A549 cells were treated with TNF-α alone or with BCG (MOI of 1: 25 and 1:50, 12 h) and TNF-α as shown. The MTT assay was performed at various time points to measure the cell viability. (F) Quantitative real time RT-PCR for p53-responsive pro-apoptotic genes in A549 cells with indicated treatment. (G) A549 cells were infected with BCG for 12 h prior to treatment with TNF-α. Representative immunofluorescence images and MFI for Annexin V-FITC staining. Data is representative of mean ± SEM of at least 3 different experiments and all blots are representative of 3 independent experiments. **p < 0.05 (one-way ANOVA), as compared to Med; *p < 0.05 (one-way ANOVA), as compared to TNF-α treated cells. Med, Medium.