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. 2020 Sep 15;18(9):e3000852. doi: 10.1371/journal.pbio.3000852

Fig 2. Division of cells with amplified centrioles.

Fig 2

(A) Maximum projection image of immunofluorescence in olfactory epithelium cryosections from an adult mouse expressing eGFP-centrin2. Dashed line marks the apical surface of the olfactory epithelium. Double solid line marks the basal lamina. Box marks the location of the inset shown in A’. Scale bar = 5 μm. Inset (A’) shows a deconvolved maximum projection of centrioles shown in panel A. eGFP-centrin2 and γ tubulin mark 2 clusters of centrioles. Phospho-H3 marks a cell in an early phase of mitosis. DAPI shows DNA condensed near the basal lamina and is excluded from the merge. Inset scale bar = 2 μm. (B) Immunofluorescence in cryosections of olfactory epithelium from a wild-type adult mouse. DAPI shows DNA condensed and aligned in metaphase with strong phospho-H3 colocalization. Centrin marks the distal ends of centrioles. In this single optical section, centriole clusters are shown at the spindle poles, marked by acetylated tubulin. DNA is excluded from the merge. Scale bar = 2 μm. (C) Single optical section image of immunofluorescence in olfactory epithelium from a wild-type adult mouse. Diffuse phospho-H3 marks a cell in anaphase. White arrows denote clusters of centrioles on opposite sides of the dividing cell. Scale bar = 2 μm. (D) Maximum projection of a deconvolved fluorescence image in olfactory epithelium from a mouse at embryonic stage E12.5. DAPI shows DNA condensed and aligned in metaphase. One spindle pole is shown with a cluster of centrioles marked by eGFP-centrin2. Scale bar = 2 μm. (E) Fluorescence image of a mouse at embryonic stage E12.5. DAPI shows DNA decondensing to form 2 daughter cell nuclei. White lines show cell boundaries, including a cleavage furrow, as approximated by cytoplasmic eGFP-centrin2 resulting from overexpression. Boxes mark the location of inset images. Scale bar = 5 μm. The insets (E’ and E”) show deconvolved maximum projection images of centriole clusters in the top and bottom boxes, respectively. Centrioles are marked by eGFP-centrin2. Inset scale bars = 2 μm. See S2 Fig for additional details. Arl13b-mCherry, ADP-ribosylation-factor-like GTPase 13b; eGFP, enhanced green fluorescent protein; phospho-H3, phosphorylated histone 3.