Fig. 3

circIPO11 overexpression promotes liver CSCs self-renewal and tumorigenic capacity. A CircIPO11 overexpressed HCC cell lines and HCC primary cells were established with lentivirus carrying indicated mini-gene constructs, followed by Northern blot. oeVec, overexpression empty vector; oecirc, overexpression of circIPO11. CircIPO11 and 18S rRNA (loading control) probes were used for Northern blot. Data are shown as means ± SD. B, C CircIPO11 overexpression enhanced the capacity of oncosphere formation. Representative images (B) and statistical results of sphere-formation ratios (C) are shown. Scale bar, 500 μm. Data are shown as means ± SD. D Diluted circIPO11 overexpression or control Huh7 CSCs were subcutaneously injected into BALB/c nude mice for 3 months to determine tumor-bearing rates of mice. Tumor-free mice are counted (left panel) and representative tumor images are shown (right panel). n = 8 for each group. E 1 × 10⁶ circIPO11-overexpressing or control HCC cells were subcutaneously injected into BALB/c nude mice. Results are shown as means ± SD. Representative tumor images are shown (right panel). n = 5 for each group. F Orthotopic liver tumor imaging of Huh7-Luc cells transduced with control or oecircIPO11 vectors. Representative pictures are shown (left panel), and statistical results are shown as means ± SD (right panel). n = 5 for each group. G Overexpression of circIPO11 enhanced the capacity of clone formation. Representative images and statistical results are shown. *P < 0.05; **P < 0.01; *** P < 0.001 by two-tailed Student’s t test. Data are representative of at least three independent experiments