Journal: bioRxiv
Article Title: Keratin 15 promotes a progenitor cell state in basal keratinocytes of skin epidermis
doi: 10.1101/2025.02.27.640633
Figure Lengend Snippet: A. Krt14 C373A/WT and Krt14 C373A/null (arrow) littermates at P5. B. H&E staining performed on back skin harvested from P5 Krt14 C373A/WT and Krt14 C373A/null littermate pups. Scale bar, 50 µm. C. H&E staining performed on tail tissue harvested from 8 week old Krt14 C373A/WT and Krt14 C373A/null tail tissue. Scale bar = 50 µm. D. Mean cytoplasmic area of basal keratinocytes with standard deviation. Dots represent single cells. Comparisons made using Mann-Whitney tests. * P = 0.0286. E. Basal cell density calculated by mean cells/µm of basal lamina with standard deviation. Dots represent individual micrographs from tail skin harvested from 4 littermate animals each. Comparisons made using Mann-Whitney tests. * P = 0.014. F. Ki67 immunofluorescence performed on 8 week old Krt14 C373A/WT and Krt14 C373A/null littermate tail skin. Merge panel displays Ki67 (green) with DAPI counterstain (red). Arrows indicate Ki67-positive basal nuclei. Dashed line specifies the basal lamina. epi, epidermis; derm, dermis. Scale bar, 50 µm. G. Mean of the Ki67 mean intensity value for individual basal keratinocyte nuclei, displayed with standard deviation. Percentages of cells above the visual threshold for Ki67 positivity (green dashed line). Comparisons made using Mann-Whitney tests. **** P <0.0001. H. Representative transmission electron microscopy micrographs of basal keratinocytes in ear skin if 8 week old Krt14 C373A/WT and Krt14 C373A/null . Nu, nucleus; kf, keratin filaments; arrows, hemidesmosomes; *, areas devoid of keratin filaments. Scale bar, 800 nm. I. Mean of the basal lamina processed/ distance covered to represent basal lamina convolution, displayed with standard deviation. Dots are individual micrograph images. Comparisons made using Mann-Whitney tests. *** P = 0.0009.
Article Snippet: Slides were blocked in 5% normal donkey serum/1% BSA/PBS for 1 h at RT, incubated in primary antibody (rabbit anti-Ki67 (Cell Signaling Technology, 9129S); chicken anti-keratin 15 (BioLegend, 833904)) for 16 h at 4 °C, Alexa Fluor-conjugated secondary antibody for 1 h at room temperature, counterstained with DAPI, and mounted in Fluorsave (Millipore, 345789) and a #1.5 coverslip.
Techniques: Staining, Standard Deviation, MANN-WHITNEY, Immunofluorescence, Transmission Assay, Electron Microscopy
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