Extended Data Fig. 3: Characterization of Keratin 10 reporter expression and basal cell delamination.
(A) Quantification of K10 reporter fluorescence in individual cells used for Fig. 1F. Dotted lines indicate the threshold used to score cells as reporter positive. (B) Percent tracked cells scored as K10 reporter positive at the timepoint prior to complete loss of ECM contact. N = 90 cells from 3 mice. Error bars are mean ±S.D. (C, D and E) Representative images and corresponding fluorescent quantification of three individual revisited basal cells as they induce K10 reporter expression and exit the basal layer. Each cell and fluorescent track were selected from the 90 cells from 3 independent mice shown in (A). Scale bar = 10 µm. (E) Lifeact-GFP fluorescence (green) in the basal layer of ear epidermis. Insets show a cell in the process of delaminating (arrows). Top inset is an xy section from the upper half of the cell; middle inset is an xy section from directly above the ECM signal (blue); bottom inset is a lateral reslice. Image is representative of data from n = 3 mice. (F) GFP-tagged Non-muscle myosin IIB (GFP-NMMIIB) (green) in the basal layer of ear epidermis. K14H2B-mCherry signal (red) shows positions of epithelial nuclei. Insets show a cell in the process of delaminating (arrows). Top inset is an xy section from the upper half of the cell, containing the cell nucleus; middle inset is an xy section from directly above the ECM signal (blue); bottom inset is a lateral reslice. Image is representative of data from n = 3 mice. Scale bars for (E) and (F) = 20 µm or 10 µm (smaller insets).
