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. 2022 Jul 25;48(3):166. doi: 10.3892/or.2022.8381

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Copyright: © Li et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 4. — Cx32 regulates the expansion of LCSCs. (A) Effect of Cx32 knockdown on the mRNA expression levels of stemness-associated genes in HepG2 cells. (B) Effect of Cx32 overexpression on the mRNA expression levels of stemness-associated genes in HCCLM3 cells. (C and D) The expression levels of stemness-associated genes were examined by western blotting when HepG2 cells were transfected with shRNA-Cx32. The proteins were normalized with β-actin. (E and F) The expression levels of stemness-associated genes were observed when HCCLM3 cells overexpressed Cx32. The proteins were normalized with β-actin. (G and H) Effect of Cx32 knockdown on the sphere-forming capacities of HepG2 cells. Magnification, ×200. The bar graph shows the average number of spheres >100 µm in diameter. (I and J) Effect of Cx32 overexpression on the sphere-forming capacities of HCCLM3 cells. Magnification, ×200. The bar graph shows the average number of spheres >100 µm in diameter. For the aforementioned images, the error bars represent the mean ± SEM of three independent experiments; *P<0.05 vs. the NC group; or *P<0.05 vs. the overexpression EV group. (K and L) Efficiency of tumor formation of Cx32 overexpression in HCCLM3 cells. (M) Efficiency of tumor formation of HCCLM3 EV and HCCLM3 OE cells; number of injected cells: 1×103 and 5×103, n=8. (N) Efficiency of tumor formation of HCCLM3 EV and HCCLM3 OE cells; number of injected cells: 1×104, 5×104, n=8. Cx32, connexin 32; LCSCs, liver cancer stem cells; shRNA, small interfering RNA; NC, negative control; EV empty vector; OE, overexpression; EpCAM, epithelial cell adhesion molecule.