FIGURE 7.

Domain swapping and substitution analysis between hZnT10 and hZnT2. A, expression of hZnT2_(H106N) failed to confer manganese resistance in SPCA1^−/−/− cells. B, expression of hZnT2_(H106N) lost the ability to confer zinc resistance in ZnT1^−/−MT^−/−ZnT4^−/− cells. C, expression of hZnT10_(hZnT2Cter) lost the ability to confer manganese resistance in SPCA1^−/−/− cells. D, expression of hZnT10_(hZnT2Cter) did not confer zinc resistance in ZnT1^−/−MT^−/−ZnT4^−/− cells. E, expression of hZnT2_(hZnT10Cter) did not confer manganese resistance in SPCA1^−/−/− cells. F, expression of hZnT2_(hZnT10Cter) lost the ability to confer zinc resistance in ZnT1^−/−MT^−/−ZnT4^−/− cells. In A, C, and E or in B, D, and F, cells were grown as presented in Fig. 3, A and B, and the numbers of living cells were evaluated by the Alamar Blue assay. A–F, Alamar Blue assay was performed at least three times. Confirmation of stable expression of WT and mutants of hZnT2 and hZnT10 in SPCA1^−/−/− cells or ZnT1^−/−MT^−/−ZnT4^−/− cells by immunoblotting (lower panels) is shown. Tubulin is shown as the loading control.