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Fig. 8. Classification of MR cells in the yolk-sac membrane of Mozambique tilapia embryos into four types, by means of quintuple-color immunofluorescence staining: type-I (A), type-II (B), type-III (C), and type-IV (D). The signals for Na+/K+-ATPase (red), NKCC1a (blue), NCC (cyan), NHE3 (yellow) and CFTR (green) are shown as separate channels, and the five channels are merged in Merged X­Y and X­Z planes. Merged X­Z plane, the X­Z optical section cut transversely at the horizontal lines indicated in Merged X­Y plane. Merged X­Y plane, the X­Y optical section cut at the lines indicated in Merged X­Z plane. n, Nucleus. Bar: 10 μm. Schematic diagrams of each of the four cell type are presented in the bottom row, showing the apical or basolateral localization patterns of Na+/K+-ATPase (red), NKCC1a (blue), NCC (cyan), NHE3 (yellow) and CFTR (green). Reprinted from J. Exp. Biol., 211, Hiroi et al., Evidence for an apical Na­Cl cotransporter involved in ion uptake in a teleost fish, 2584–2599, 2008, The Company of Biologists Limited.

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