Figure 4.

In clonal HepG2 cells, overexpression of SLC30A10-WT or SLC30A10-T95I has a comparable effect on intracellular Mn levels and Mn-induced cell death. A: immunoblot analyses were performed to detect SLC30A10 (using the custom anti-SLC30A10 antibody) or tubulin in HepG2 clones expressing SLC30A10-WT or SLC30A10-T95I. Relative expression of SLC30A10-WT or T95I, normalized to tubulin, respectively is 1.00 ± 0.136 and 0.541 ± 0.021 (means ± SE; n = 3, P < 0.05 by t test). Intracellular Mn (B), Fe (C), Cu (D), or Zn (E) levels were assayed in mock-infected HepG2 cells (control) or HepG2 clones expressing SLC30A10-WT or SLC30A10-T95I after 16 h treatment with 125 µM Mn. Metal values were normalized to total cell counts (means ± SE; n = 7–8, *P < 0.05 and n.s. denotes not significant for indicated comparisons by one-way ANOVA and Tukey’s post hoc test). F: viability of cells infected as described in B–E was assayed after 16 h treatment with indicated Mn doses. For each infection condition, viability at 0 mM Mn was set to 100 (means ± SE; n = 5, *P < 0.05 using two-way ANOVA and Tukey’s post hoc test with a indicating differences in comparison with the control group at each Mn dose. There were no differences between SLC30A10-WT or SLC30A10-T95I infection groups at any of the Mn doses by two-way ANOVA (P > 0.05). WT, wild type.