Extended Data Fig. 3: Phenotypic consequence of an HFSC specific deficiency of the aPKCλ gene. | Nature Aging

Extended Data Fig. 3: Phenotypic consequence of an HFSC specific deficiency of the aPKCλ gene.

From: Distinct types of stem cell divisions determine organ regeneration and aging in hair follicles

Extended Data Fig. 3

a, Representative IF images of KRT15 and aPKCλ in the bulge area and epidermis of control and aPKCλ cKO mice at 11 weeks of age. b, Quantification of the percent of aPKCλ deficient HFs in control and in aPKCλ cKO mice (control, n = 14 images; aPKCλ cKO, n = 14 images). aPKCλ cKO mice have a significantly decreased expression of aPKCλ in the bulge area. c, Quantification of percent cells with polarized localization in the Bg and sBg areas of Control (n = 9 mice, n = 43 cells), aPKCλ cKO (n = 4 mice, n = 25 cells) mice. aPKCλ deficiency significantly abolished the polarized localization. d, IF images of KRT15 and KRT1 in the dorsal skin of 11 wo control and aPKCλ cKO mice at 3 days after HC induction. e, Quantification of the percentage of KRT1+ cells per each HF after HC induction (control, n = 34 images; aPKCλ cKO, n = 8 images). aPKCλ cKO significantly increased the percentage of KRT1+ cells per each HF. f, IF images of KRT1 and KRT75 in control or in aPKCλ cKO mice at 3 days after HC induction. aPKCλ deficient-HFSCs differentiate into KRT1+ cells through SR type of ACDs. g, IF images of LacZ and LRIG1 in the dorsal skin of 11 wo control and aPKCλ cKO mice at 3 days after HC induction. LacZ+LRIG1+ (arrows) in HFSCs were found in aPKCλ cKO mice. h, Gene set enrichment analysis (GSEA) profile with ‘RAMALHO STEMNESS UP’ molecular signature in anagen stage (aHFSC) fraction between Cont and aPKCλ cKO mice. i, Heat map showing the log fold expression changes (logFC) of HFSC signature genes between RU486-treated control (Cont) and aPKCλ cKO telogen stage HFSCs (tHFSCs) from microarray data. aPKCλ cKO tHFSCs had decreased expression of HFSC signature genes excluding the Col17a1 gene. j, Heat map showing the log fold change of epidermal differentiation-related genes between Cont and aPKCλ cKO aHFSCs from microarray data. aPKCλ cKO aHFSCs had increased expression of epidermal differentiation-related genes including NOTCH related genes. k, IF images of NOTCH1, KRT1 and KRT15 in 11 wo Cont and aPKCλ cKO mice. NOTCH1+KRT1+ (arrows) in KRT15+ HFSCs were found in 11 wo aPKCλ cKO mice at 3 days after HC induction. l, IF images of c-MYC and KRT15 in 11 wo Cont and aPKCλ cKO mice. c-MYC+ cells (arrows) in KRT15+ HFSCs were found in 11 wo aPKCλ cKO mice at 3 days after HC induction. m, IH images of LacZ and Oil red O staining in 15 wo Cont and aPKCλ cKO mice. HFSC-derived LacZ+ cells (arrows) migrated into the epidermis together with SGs in 15 wo aPKCλ cKO mice at 1 month after HC induction. n, Schematic for experimental hypothesis of the interaction between the loss of aPKCλ and hemidesmosomes. o, p, q, Representative IF images of KRT15, COL17A1 (o), ITGA6 (p) and ITGB4 (q) in the bulge area of control and aPKCλ cKO mice at 11 weeks old. A deficiency of aPKCλ did not affect the expression level of HD components, COL17A1, ITGA6 and ITGB4. Error bars, means ± SEM; White dashed line, basement membrane; white line and fan-shaped line and upper number, spindle angle against basal cell layer (o); green(control)/red(cKOs) dashed line, dividing HFSCs. Chi-square test (c) or two-tailed Student’s t test (b, e).

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