Fig. 3

SPAG5 promotes HCC proliferation in vitro and in vivo. a and b, The efficiency of SPAG5 silencing in HCC cell lines was measured by RT-PCR and Western blot. c, The proliferation capacities were detected by CCK8 assays in Huh7 and HCCLM3 cells transfected with the shNC or the shSPAG5 plasmid ^*p < 0.05. d, EdU results showing the effect of SPAG5 knockdown on the proliferation abilities of Huh7 and HCCLM3 cells, ^*p < 0.05. e and f, Detection for cell cycle of HCC cells after silencing SPAG5 expression in Huh7 and HCCLM3 cells. Results are expressed as peak diagram (left) and calculated distribution for cells in G0/G1, S, and G2/M phases (right). ^*p < 0.05. g and h, Measurement of apoptotic cells under SPAG5 down-regulation in Huh7 (g) and HCCLM3 cells (h). Results are expressed as scatter diagram (left) and calculated percentage of annexin-V-positive cell population (right). ^*p < 0.05. i and j, The in vivo tumor formation was examined by subcutaneously injecting shNC or shSPAG5 cells into the flank of nude mice (n = 6), ^*p < 0.05. k and l, representative examples of the immunohistochemistry analyses of xenograft tumors with anti-C-caspase-3 antibodies. Scale bar, 50 μm. Quantifcation of anti-C-caspase-3–positive cells of three individual tumors. ^*p < 0.05