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. 2023 Apr 7;9(14):eadd5745. doi: 10.1126/sciadv.add5745

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Fig. 6. — (A) In situ HCR and validation of lineage inference over 10 development stages of the embryonic epidermis in embryos, marking ionocytes (atp6v1b1.L; orange), multiciliated (tekt2.S; green), and basal cells (pfn1.L; magenta). The nuclei are marked by 4′,6-diamidino-2-phenylindole (DAPI) staining (cyan). The multiciliated cells and ionocytes emerge during neurula in the sensorial layer and radially intercalate (move apically) to superficial layers by early tailbud stages. Multiciliated cells are consistently observed at stage 13 (stage 13A), whereas ionocytes are consistently observed at stage 16; in four of nine embryos, ionocytes are observed at stage 13 (stage 13B). The basal cells are retained in the sensorial layer. The yellow dashed rectangles indicate the zoomed-in regions shown in (B). Yellow arrowheads point to regions (within the nucleus) expressing tekt2.S. (B) Zoomed view of in situ HCR marking expression, positioning, and migration of multiciliated (tekt2.S; green), ionocytes (atp6v1b1.L; orange), and basal cells (pfn1.L; magenta). Images in (A) and (B) represent the maximum intensity projections of transverse cross sections, and white dashed lines indicate the apical superficial epithelium. (C) In situ HCR of whole embryonic surface epithelium marked by multiciliated (tekt2.S; green) and ionocytes (atp6v1b1.L; orange) at the superficial epidermis. Multiciliated cells and ionocytes move apically (radially intercalate) from the sensorial to the superficial epithelium and distribute in a salt-and-pepper fashion. Images represent the maximum intensity projections of Z sections taken from the apical superficial epithelium. The dashed white rectangles indicate the zoomed-in regions shown in (C), bottom row.