Figure 4. NUDT1 silencing decreases survival, migration, and invasion of HCC cells. (A) Western blot analysis shows NUDT1 expression in HCC cell lines, Hep-3B, SK-Hep-1, PLC, BEL-7402 and MHCC-97H, and the hepatic immortalized cell line, LO2. The relative levels of endogenous NUDT1 expression are shown in the right panel. (B) Western blot analysis shows NUDT1 protein levels in BEL-7402 cells that are transfected with three NUDT1-specific shRNAs (sh-NUDT1_1, sh-NUDT1_2, and sh-NUDT1_3) and negative control shRNA (sh-NC). The relative levels of NUDT1 protein are shown in the right panel. (C) Representative images show the wound healing assay results in control and NUDT1-silenced BEL-7402 at 0 and 48 h after scratching (Scale bars:100μm). The results show that NUDT1 silencing inhibits migration of HCC cells. (D) CCK8 assay results show that NUDT1 silencing decreases the viability of BEL-7402 cells compared to controls. (E) Representative images show Transwell migration and invasion assay results of control and NUDT-silenced BEL-7402 cells (Scale bars: 100μm). As shown NUDT1 silencing decreases the invasion and migration of BEL-7402 cells. (F) Colony formation assay results show the total number of colonies in control and NUDT1-silenced BEL-7402 cells. As shown, NUDT1 silencing decreases the colony formation in BEL-7402 cells. All the values are shown as mean ±SD of three independent experiments. Note: *** denotes P<0.001 as evaluated by the Student’s t-test.