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. 2021 Dec 6;10:e75050. doi: 10.7554/eLife.75050

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© 2021, Seleit et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 3. — (A–A’’) (eGFP-cbx1b) (mScarlet-pcna) double positive stage 40 medaka embryo. Maximum projection of a mature secondary neuromast (centre of image) within the lateral line system surrounded by epithelial cells, labelled by: endogenous eGFP-Cbx1b in (A) and endogenous mScarlet-Pcna in (A’). The merge is shown in (A’’). (A) eGFP-Cbx1b is a ubiquitous nuclear marker and labels all cell types within a mature neuromast. Those are: hair cells (HCs) in the centre of a neuromast, surrounded by support cells (SCs), and an outer ring of mantle cells (MCs) surrounded by the elongated neuromast border cells (nBCs). (A’) mScarlet-Pcna labels cycling cells, which are located at the very edge of the mature neuromast organ, a proportion of MCs express mScarlet-Pcna (white arrowheads). nBCs also express mScarlet-Pcna. Speckles can be seen in several mScarlet-Pcna-positive nBC nuclei (yellow arrowheads), indicating cells in late S phase of the cell cycle. (A’’) Merged image. n = 10 neuromast organs. NU = neuromast. Scale bar = 20 µm. (B–B’’) (eGFP-cbx1b) (mScarlet-pcna) stage 40 medaka embryo. Single Z-slice showing the medaka optic tectum. (B) eGFP-Cbx1b is a ubiquitous nuclear marker, (B’) whereas mScarlet-Pcna labels a subset of cells at the outer periphery of the optic tectum, indicating the position of proliferative cells in this tissue. A graded expression of mScarlet-Pcna is observed, with more central cells in the optic tectum losing the expression of mScarlet-Pcna. (B’’) Merged image. n = 4 embryos. OT = optic tectum. Scale bar = 30 µm. (C–C’’) Maximum projection of the pectoral fin of stage 40 medaka embryos. (C) eGFP-Cbx1b is a ubiquitous nuclear marker, (C’) while a subset of cells is labelled by mScarlet-Pcna indicating the position of proliferative cells. Note the proximal to distal gradient of mScarlet-Pcna expression, with proliferative cells at the base of the fin (left) and differentiated cells at the outer edges of the fin (right) (C’’) Merged image. n = 4 embryos. PC = pectoral fin. Scale bar = 50 µm.

Figure 3—figure supplement 1. — (A–A’’) (eGFP-cbx1b) (mScarlet-pcna) double positive stage 40 medaka embryo. Maximum projection of a mature secondary neuromast (centre of image) within the lateral line system surrounded by epithelial cells, labelled by: endogenous eGFP-Cbx1b in (A) and endogenous mScarlet-Pcna in (A’). The merge is shown in (A’’). (A) eGFP-Cbx1b is a ubiquitous nuclear marker and labels all cell types within a mature neuromast. Those are: hair cells (HCs) in the centre of a neuromast, surrounded by support cells (SCs), and an outer ring of mantle cells (MCs) surrounded by the elongated neuromast border cells (nBCs). (A’) mScarlet-Pcna labels cycling cells, which are located at the very edge of the mature neuromast organ, a proportion of MCs express mScarlet-Pcna (white arrowheads). nBCs also express mScarlet-Pcna. Speckles can be seen in several mScarlet-Pcna-positive nBC nuclei (yellow arrowheads), indicating cells in late S phase of the cell cycle. (A’’) Merged image. n = 10 neuromast organs. NU = neuromast. Scale bar = 20 µm. (B–B’’) (eGFP-cbx1b) (mScarlet-pcna) stage 40 medaka embryo. Single Z-slice showing the medaka optic tectum. (B) eGFP-Cbx1b is a ubiquitous nuclear marker, (B’) whereas mScarlet-Pcna labels a subset of cells at the outer periphery of the optic tectum, indicating the position of proliferative cells in this tissue. A graded expression of mScarlet-Pcna is observed, with more central cells in the optic tectum losing the expression of mScarlet-Pcna. (B’’) Merged image. n = 4 embryos. OT = optic tectum. Scale bar = 30 µm. (C–C’’) Maximum projection of the pectoral fin of stage 40 medaka embryos. (C) eGFP-Cbx1b is a ubiquitous nuclear marker, (C’) while a subset of cells is labelled by mScarlet-Pcna indicating the position of proliferative cells. Note the proximal to distal gradient of mScarlet-Pcna expression, with proliferative cells at the base of the fin (left) and differentiated cells at the outer edges of the fin (right) (C’’) Merged image. n = 4 embryos. PC = pectoral fin. Scale bar = 50 µm.