Figure 4.

Knockdown of YB-1 does not inhibit proliferation, but suppresses migration, invasion, and sphere formation of HuH-7^R cells. (A) HuH-7^R cells were infected with lentiviruses containing shYB-1 (#1, #2) or shCtrl, and after 48 h, lysed and analyzed for YB-1 expression status by immunoblotting with the indicated antibodies. (B) Viability of HuH-7^R cells treated with shCtrl, shYB-1 or 5 μM sorafenib determined at the indicated time-points with the MTT ssay. Plots depict cumulative cell absorbance versus days in culture. (C) Wound healing assay of YB-1 knockdown HuH-7^R cells. The micrographs depict cells migrating into the gap 0 h and 15 h after removal of the insert. (D) Transwell invasion assay of YB-1 knockdown HuH-7^R cells. Cells in the central field of each insert were visualized using light microscopy and quantified. (E) Sphere formation assay of YB-1 knockdown HuH-7^R cells. Images of the spheres formed were captured on day 9 and sphere diameters measured. Data were quantified and presented as means ± SD. All results are the representatives of at least three independent biological replicates. Scale bars = 50 µm. p Values were determined using the t test (ns, not significant; ** p < 0.01); shCtrl, control shRNA; shYB-1, shRNA against YB-1.