Figure 8

(a) Silencing of TPD52 in CWR22Rν1 cells. Western blot analysis of TPD52 protein expression in siRNA-TPD52 transfected CWR22Rν1 cells. The total cell lysate was prepared and 40 μg protein was subjected to SDS-page followed by Immunoblot analysis and chemiluminescence detection. The blots shown here are representative of three independent experiments with similar results. (b) Effect of downregulation of TPD52 on the proliferation rate of 22Rv1 cells. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assays were performed after downregulation of TPD52 in 22Rv1 cells. Downregulation of TPD52 leads to decreased cell proliferation as compared to control untransfected and scrambled RNA transfected cells. (c) Immunohistochemistry of TPD52-siRNA down-regulated CWR22Rν1 cells. As TPD52 is localized mainly in the cytoplasm so brown (DAB) stain shows an expression of TPD52 and the blue counterstain (hematoxylin) represents the nucleus. The first left represents control untransfected CWR22Rν1 cells showing high staining of TPD52, the middle is scrambled RNA transfected cells also show staining close to normal cells and the third is SiRNA-TPD52 transfected CWR22Rν1 cells showing low to mild staining of TPD52. (d) Migration of TPD52-siRNA down-regulated CWR22Rν1 cells. As shown by microscopic analysis, the first left figure represents untransfected normal CWR22Rν1 cells showing almost 60% of migrated cells, the middle figure represents scrambled RNA transfected cells showing almost the same number of migrated cells as that of normal control and third is SiRNA-TPD52 transfected CWR22Rν1 cells which show only 10% of migrating cells.