Figure 3.

DDX59 is important to maintain cancer cell phenotype. (a) Calu-1, SK-LU-1, HCC827, and H1299 lung cancer cells were either infected with shSCR or shRNA-DDX59 lentivirus. Four days post lentiviral infection, cells were collected. Whole-cell lysates were subjected to western blot analysis with anti-DDX59 and anti-tubulin antibodies. (b) Three days post infection, cells were split and ~5000 cells per condition were plated on 100-mm dish in triplicates. Cells were further cultured for 2 weeks until being subjected to Foci analysis. Typical images for the culture of the indicated transduced cells are shown. (c) SK-LU-1, HCC827, and H1299 cells were transduced with either shSCR or shRNA-DDX59 lentivirus. Cells were subjected to soft-agar analysis after plating 1×10⁴ cells into each 60-mm dish. Typical image is shown after culturing for 2 weeks. (d) Quantitation of colony numbers in the soft-agar assay is graphed; error bars were taken from triplicates. *P<0.05, n=3. (e) Above-mentioned cells were subjected to flow cytometry analysis after staining with Annexin V and PI using the Vybrant apoptosis kit. Percentage of apoptotic cells were labeled in the quadrants.