Figure 3

DDX21 knockdown in multiple breast cancer cell lines induces cell death and cell cycle arrest. (A) MCF-7 and T47D breast cancer cells were either infected with shSCR or shRNA-DDX21 lentiviruses. Three days postinfection, cells were split and cultured for 48 hours. Typical images for the live culture of the indicated transduced cells are shown. (B) MDA-MB-231 cells or HCC1806 cells were either transduced with shSCR or shRNA-DDX21 lentiviruses. Whole cell lysates were subjected to western blot analysis four days and five days postinfection with anti-DDX21 and anti-tubulin antibodies. (C) MDA-MB-231 cells were either transduced with shSCR or shRNA-DDX21 lentiviruses. Five days postinfection, cells were then subjected to apoptosis analysis by flow cytometry with Vybrant apoptosis kit after PI labeling of nuclei and FITC labeling of Annexin V. Percentage of live and dead cells are shown in each quadrant. (D) Quantitation of live and apoptotic cells were calculated and graphed. (E) Above-mentioned MDA-MB-231 cells were subjected to fixation and PI staining before flow cytometry to detect cell cycle distribution. Percentage of cells in each cell cycle phase is presented. (F) HCC1806 cells were either transduced with shSCR or shRNA-DDX21 lentiviruses. Five days postinfection, cells were subjected to apoptosis analysis. Population of live and dead cells is marked in each quadrant. (G) Quantitation of the live and apoptotic cells is graphed. FITC, fluorescein isothiocyanate.