Figure 5.

ID enhanced HCC cell migration and invasion in vitro

(A–D) Huh7 cells or primary hepatocytes were treated with either dimethyl sulfoxide (DMSO) or iron chelators, deferoxamine (DFO) and Dp44mT, for 24 h. (A) The number of dividing Huh7 cells were determined by EdU fluorescence levels. (B) Migration and invasion ability of Huh7 cells under ID. (C and D) The mRNA (left) and protein (right) expression of SPNS2 and iron-related genes in the (C) Huh7 cells and (D) primary human and mouse hepatocytes under ID. See experiment results using other HCC cell lines in Figures S8–S10. (E) Efficiency of siRNA-mediated SPNS2 knockdown in the Huh7 cells. (F) Huh7 cells were transfected with siRNA against SPSN2 and co-treated with either DMSO or DFO for 24 h. Migration and invasion abilities of SPNS2 knockdown Huh7 cells under ID. Representative figures were shown. See experiment results using other HCC cell lines in Figure S12. All in vitro experiments were performed as 3 replications. Both representative figures and quantitative data of western blot assays were shown. Data were presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01 versus DMSO, or versus siNC. Edu, 5-ethynyl-2′-deoxyuridine.