Fig. 2

CircUCK2(2,3) promotes HCC cell proliferation and metastasis both in vitro and in vivo. A and B CCK8 assays (A) or colony formation assays (B) in SNU182 cells with circUCK2(2,3) knockdown, (scale bar, 5 mm). C EdU staining assays in SNU182 cells with circUCK2(2,3) knockdown, (scale bar, 40 μm). D Transwell migration and invasion assays in SNU182 cells with circUCK2(2,3) knockdown, (scale bar, 40 μm). E Western blotting and densitometry of EMT markers in SNU182 cells with circUCK2(2,3) knockdown. F–H PLC/PRF/5 cells with stable circUCK2(2,3) knockdown were injected into nude mice subcutaneously. Seven days after injection, tumor volumes were measured three times weekly by using a caliper to draw tumor growth curve (F) (means ± SD, two-way ANOVA). After 23 days of injection, subcutaneous tumors were dissected and imaged (G). Tumor weight was compared between the sh-NC group and the groups with circUCK2(2,3) knockdown (H). I IHC staining of Ki67 in xenograft tumors (scale bar, 100 μm). J PLC/PRF/5 cells with stable circUCK2(2,3) knockdown were introduced into nude mice intravenously. Eight weeks after the initial injection, lungs were dissected and lung micrometastases were detected by H&E staining (scale bar, 1 mm and 200 μm). Statistical analyses were performed using paired Student’s t-tests (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). siNC negative control of siRNA, si-1 siRNA#1, si-2 siRNA#2, VC vector control, OE overexpression, DAPI 4′,6-diamidino-2-phenylindole, EdU 5-ethynyl-2′-deoxyuridine, sh-NC negative control of shRNA, sh-1 shRNA#1, sh-2 shRNA#2, Ki67 marker of proliferation