Figure 3. p-NPM-Thr^234/237 enhanced HCC cell migration and invasion.

A. Endogenous NPM was knocked down by using lentiviral based knockdown approach. By western blot, NPM protein was successfully repressed in Hep3B and Huh7 cells. B. NPM-wild type (NPM WT) and NPM Thr234/237A mutant were efficiently overexpressed in Hep3B and Huh7 cells with a lentiviral-based approach, confirmed by protein expression of Flag observed in NPM WT and NPM Thr234/237A transfectants. C. BrdU cell proliferation assay demonstrated that NPM wild-type Hep3B and Huh7 cells (NPM WT) had significantly increased cell proliferation rate when compared with control (EV); whereas the proliferative effect is partially abolished in NPM Thr234/237A transfectants (*p < 0.05, **p < 0.01 and ***p < 0.001, student t test). D. Cell Migration and E. invasion assays demonstrated that NPM Thr234/237A mutant transfected Hep3B and Huh7 cells (NPM Thr234/237A) showed reduced migratory and invasion abilities when compared with respective NPM wild-type transfectants (NPM WT) (**p < 0.01 and ***p < 0.001, student t test) (magnification × 10).