Supplementary Figures 1 - 3, Table 1 from SIRT1 Inactivation Evokes Antitumor Activities in NSCLC through the Tumor Suppressor p27

Fig.S1: The level of p27Kip1 mRNA was unaffected by SIRT1 silencing. The mRNA levels of p27 were measured by quantitative RT-PCR analysis. 5 mg RNA extracted from SIRT1 silenced (shSIRT1) or shRNA control (shControl) H1299 and H460 cells. The mRNA levels of p27 are expressed relative to b-actin transcripts. Each experiment was performed in triplicate and repeated three times. The error bars represent the SEM. Fig.S2: SIRT1 silencing has no effect on apoptosis. Cell extracts were made from SIRT1- silenced and shRNA-control H1299 and H460 cells, and immunoblot analysis was performed with PARP and β-actin antibodies. Fig.S3: A. SIRT1 has no effect on p21 expression in both p53 wild type and p53 null cells. Cell extracts were made from SIRT1-silenced and shRNA-control H460 (p53+/+) and H1299 (p53-/-) cells, and immunoblot analysis was performed with anti-acetylated-p53, p21 and β-actin antibodies. B. p16 is deleted in NSCLC H460 and A549 cells. Cell extracts were made from SIRT1-silenced and shRNA-control H460 (p16-/-) and A549 (p16-/-) cells, or p16 wild type Hela cells. The immunoblot analysis was performed with anti-acetylated-p16, and β-actin antibodies. Table 1. The p27, p53 and p16 status in studied NSCLC cell lines.