Fig 1: Expression of sulfatide species with phytosphingosine (t18:0) and 2-hFAs and the genes of corresponding enzymes in intercalated cells. Cells collected from collagenase-treated kidney tissues were labeled with anti-ATP6V0A4 antibodies, and the ATP6V0A4 high population was collected using a FACS Aria flow cytometer (A-F). Fractionated ATP6V0A4-positive doublet cells (B, D) were removed using FSC-W, FSC-H (E) and SSC-W, SSC-H (F). The sulfatide species in methanol extracts of the sorted ATP6V0A4-high population were analyzed using iMScope (G). Relative ratios of ion intensity for each sulfatide species in the ATP6V0A4-high population to that of the ATP6V0A4-low population, normalized to that of the PI ion (m/z 885.50), are shown (G). The ratio of m/z 896.61 and 924.64 ions in this ATP6V0A4-high population were increased in three independent experiments. The expression levels of Dsgs2 (encoding sphingolipid delta(4)-desaturase) and Fa2h (encoding FA 2-hydroxylase) in the ATP6V0A4-high population were analyzed using RT-qPCR (H). Atp6v0a4 (encoding ATP6V0A4 in intercalated cells), Slc26a4 (encoding pendrin in type B-intercalated cells), and AE1 (encoding SLC4A1 in type A intercalated cells) were used as positive controls. B2m (encoding b2-microglobulin) and Gapdh (encoding glyceraldehyde-3-phosphate dehydrogenase) were used as housekeeping genes.