Figure 7.
K15 overrepresentation alters defining attributes of basal layer keratinocytes in vivo. (A)Krt14C373A/WT and Krt14C373A/null (arrow) littermates at P5. (B) H&E staining performed on back skin harvested from P5 Krt14C373A/WT and Krt14C373A/null littermate pups. Scale bar, 50 µm. (B′) Magnified insets of H&E-stained back skin in Fig. 5 A. Scale bar, 10 µm. (C) Epidermal thickness (µm) and hair follicle depth (µm) in back skin harvested from P5 Krt14C373A/WT and Krt14C373A/null littermate pups. Comparisons were made using Mann–Whitney tests. ****P < 0.0001. (C′) Mean cytoplasmic area of basal keratinocytes with standard deviation; dots represent scs. Comparisons were made using Mann–Whitney tests. *P = 0.0286. Basal cell density was calculated by mean cells/µm of basal lamina with standard deviation; dots represent individual micrographs from tail skin harvested from four littermate animals each. Comparisons were made using Mann–Whitney tests. *P = 0.014. (D) H&E staining performed on tail tissue harvested from 8-wk-old Krt14C373A/WT and Krt14C373A/null tail tissue. Scale bar = 50 µm. (E) YAP1 and K15 immunofluorescence performed on 8-wk-old Krt14C373A/WT and Krt14C373A/null littermate tail skin. Merge panel displays DAPI counterstain (blue), YAP1 (green), and K15 (red). Arrows emphasize nuclei of suprabasal keratinocytes, showing YAP1-negative nuclei in Krt14C373A/WT tail skin and YAP1-positive nuclei in Krt14C373A/null tail skin. A dashed line specifies the basal lamina. epi, epidermis; derm, dermis. Scale bar, 50 µm. (F) Representative transmission electron microscopy micrographs of 8-wk-old Krt14C373A/WT and Krt14C373A/null basal keratinocytes in ear skin. Nu, nucleus; kf, keratin filaments; arrows, hemidesmosomes; *, areas devoid of keratin filaments. Scale bar, 800 nm. (F) Ki67 immunofluorescence performed on 8-wk-old Krt14C373A/WT and Krt14C373A/null littermate tail skin. Merge panel displays Ki67 (green) with DAPI counterstain (red). Arrows indicate Ki67-positive basal nuclei. A dashed line specifies the basal lamina. epi, epidermis; derm, dermis. Scale bar, 50 µm. (G) Mean of the basal lamina processed/distance covered to represent basal lamina convolution, displayed with standard deviation. Dots are individual micrograph images. Comparisons were made using Mann–Whitney tests. ***P = 0.0009. (G′–H′) Mean of the Ki67 MIV for individual basal keratinocyte nuclei, displayed with standard deviation. Percentages of cells above the visual threshold for Ki67 positivity (green dashed line). Comparisons were made using Mann–Whitney tests. ****P < 0.0001.

K15 overrepresentation alters defining attributes of basal layer keratinocytes in vivo. (A) Krt14 C373A/WT and Krt14C373A/null (arrow) littermates at P5. (B) H&E staining performed on back skin harvested from P5 Krt14C373A/WT and Krt14C373A/null littermate pups. Scale bar, 50 µm. (B′) Magnified insets of H&E-stained back skin in Fig. 5 A. Scale bar, 10 µm. (C) Epidermal thickness (µm) and hair follicle depth (µm) in back skin harvested from P5 Krt14C373A/WT and Krt14C373A/null littermate pups. Comparisons were made using Mann–Whitney tests. ****P < 0.0001. (C′) Mean cytoplasmic area of basal keratinocytes with standard deviation; dots represent scs. Comparisons were made using Mann–Whitney tests. *P = 0.0286. Basal cell density was calculated by mean cells/µm of basal lamina with standard deviation; dots represent individual micrographs from tail skin harvested from four littermate animals each. Comparisons were made using Mann–Whitney tests. *P = 0.014. (D) H&E staining performed on tail tissue harvested from 8-wk-old Krt14C373A/WT and Krt14C373A/null tail tissue. Scale bar = 50 µm. (E) YAP1 and K15 immunofluorescence performed on 8-wk-old Krt14C373A/WT and Krt14C373A/null littermate tail skin. Merge panel displays DAPI counterstain (blue), YAP1 (green), and K15 (red). Arrows emphasize nuclei of suprabasal keratinocytes, showing YAP1-negative nuclei in Krt14C373A/WT tail skin and YAP1-positive nuclei in Krt14C373A/null tail skin. A dashed line specifies the basal lamina. epi, epidermis; derm, dermis. Scale bar, 50 µm. (F) Representative transmission electron microscopy micrographs of 8-wk-old Krt14C373A/WT and Krt14C373A/null basal keratinocytes in ear skin. Nu, nucleus; kf, keratin filaments; arrows, hemidesmosomes; *, areas devoid of keratin filaments. Scale bar, 800 nm. (F) Ki67 immunofluorescence performed on 8-wk-old Krt14C373A/WT and Krt14C373A/null littermate tail skin. Merge panel displays Ki67 (green) with DAPI counterstain (red). Arrows indicate Ki67-positive basal nuclei. A dashed line specifies the basal lamina. epi, epidermis; derm, dermis. Scale bar, 50 µm. (G) Mean of the basal lamina processed/distance covered to represent basal lamina convolution, displayed with standard deviation. Dots are individual micrograph images. Comparisons were made using Mann–Whitney tests. ***P = 0.0009. (G′–H′) Mean of the Ki67 MIV for individual basal keratinocyte nuclei, displayed with standard deviation. Percentages of cells above the visual threshold for Ki67 positivity (green dashed line). Comparisons were made using Mann–Whitney tests. ****P < 0.0001.

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