Fig. 3: Knockdown of ZIP8 impairs ESCC cell proliferation.

A Western blot analysis confirmed the efficiency of ZIP8 knockdown in ESCC cell lines. B CCK-8 assays was used to detect the proliferation of KYSE30, KYSE450, and KYSE510 cells. C, D Colony formation assays assess the ability of KYSE30, KYSE510, and KYSE450 cells to survive and form colonies following ZIP8 knockdown. E, F Anchorage-independent cell growth assays evaluate the effects of ZIP8 knockdown on cell growth (scale bar, 200 μm). G Western blot to check the overexpression of ZIP8 in KYSE70 cells. H CCK8 assays to detect the cell proliferation after overexpression ZIP8. I–L Colony formation assays and anchorage-independent growth assays were conducted to evaluate the effect of ZIP8 overexpression on the proliferative capacity of KYSE70 cells. Representative images are shown in the upper panel, while statistical analysis is presented in the lower panel. The Student’s unpaired t-test was utilized to evaluate statistical analysis in (B, D, F, H, J, L, with *p < 0.05, **p < 0.01, p < 0.001* (n = 3, independent experiments). Data are presented as mean ± SD.