Fig. 3

TPX2 abrogation inhibits Hh signaling-induced HCC cell proliferation. a-b. HEK293 cells were transfected with shRNA-Control or three shRNA-TPX2 constructs for 48 h and were harvested for WB analysis with the indicated antibodies (a) and real-time PCR analysis with the indicated primers (b). c. The most efficient shRNA sequence for knocking down TPX2 was selected to construct lentivirus systems. Validation of Huh7 sh-TPX2 stable cell lines using WB analysis (left panel) and quantitative real-time PCR (right panel). d. TPX2 abrogation inhibited Hh signaling-induced HCC cell proliferation as determined using EdU staining. Scale bar, 100 μm. e. The ratio of EdU-positive cells was quantified using the ImageJ software (n = 3). f. Cell growth curves of sh-Control / Shh (−), sh-Control / Shh (+), sh-TPX2 / Shh (−), and sh-TPX2 / Shh (+) in Huh7 cells. g-j. Comparison of the proliferative ability of sh-Control / Shh(−), sh-Control / Shh (+), sh-TPX2 / Shh(−), and sh-TPX2 / Shh (+) in Huh7 cells using soft-agar colony formation assays (g) and plate colony formation assay (i). Soft-agar colonies (h) and plate colonies (j) were counted using the ImageJ software. Scale bar, 100 μm. Data was shown as mean ± SD (n = 3). *, p < 0.05; **, p < 0.01, N.S. denotes not significant