CNNM2- or CNNM4-transfected HEK293 cells or HEK293-derived cell lines stably expressing TpCorC were cultured in growth medium supplemented with 40 mM Mg2+ until use. The expression of each construct of TpCorC and CNNM proteins was confirmed by Western blotting (fig. S6). The Mg2+ export assay with Magnesium Green was performed as described in a previous study (15). The data are shown as either line graphs indicating the time course of mean relative fluorescent intensities or bar graphs of mean relative fluorescent intensities after Mg2+ depletion (at 5 min). Relative fluorescent intensities were calculated as the ratio of fluorescent intensity at a desired time point to that at time zero. After imaging analyses, cells were fixed with phosphate-buffered saline containing 3.7% formaldehyde and subjected to immunofluorescence microscopy to confirm protein expression. Representative line graphs are shown in the respective figures or in fig. S8.

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