Extended Data Fig. 7: Characterization of Keratin 10-positive cell divisions.
(A) UMAP representation of cell cycle phases of Joost 2020 cells that were classified as Krt10+. (B) Quantification of the proportion of S and G2/M phase cells within the Krt10+ cell population shown in (A); graph represents average of independent experiments from n = 5 mice. (C) UMAP representation of Krt10-expressing cells from the 5 biological replicates from Joost 2020, overlaid on the combined UMAP (in gray). (D) Whole mount staining of pH3 (green) and K10 (red) in the basal layer of ear skin, showing both pH3-positive, K10 negative (yellow arrows) and pH3-positive, K10-positive (white arrows) cells. Scale bar = 20 µm. (E) Whole mount staining of mVenus-hGem signal from Fucci2 reporter mice (green), K10 (red) and phalloidin (white) in the basal layer of ear skin, showing both mVenus-hGem-positive, K10 negative (yellow arrows) and mVenus-hGem-positive, K10-positive (white arrows) cells. Scale bar=20 µm. (F) Whole mount staining of EdU (red) and K10 (green) in the basal layer of dorsal skin, showing both EdU-positive, K10 negative (yellow arrows) and EdU-positive, K10-positive (white arrows) cells. Scale bar=20 µm. For (D-F), staining is representative of n = 3 mice. (G) Proportion of EdU positive cells that also express K10 protein in dorsal skin. Graph represents average from one independent immunostaining experiment using n=3 mice. (H) Proportion of divisions performed by K10 reporter positive cells in revisited basal regions. Graph represents average of n=3 imaged mice. (I) Proportion of K10 reporter positive cells performing either division or delamination as their first behavior. Analysis was limited to cells that could be seen inducing K10 reporter expression and then performing a subsequent behavior (either division or delamination) within the 6-day revisit window. Graph represents average of n=3 imaged mice. (J) Revisited K10-positive basal cell as it divides to produce one delaminating daughter cell (number 1) and a second daughter cell (number 2) that goes through another round of cell division before exiting the basal layer (numbers 2a and 2b). Scale bar = 10 µm. Image representative of data from n = 3 mice. (K) Fates of all daughter cells from Fig. 3G whose first behavior could be resolved within 5 days of birth. N = 266 cells from 3 mice. For (B), (G), (H) and (I), error bars are mean ±S.D.
