Extended Data Fig. 3: HFD feeding causes HFSCs to prematurely commit to epidermal differentiation.

a, The locations of GFP⁺ cells in K15CrePR;Rosa-H2BGFP mice after recombination (n = 2). K15CrePR:Rosa-H2BGFP mice were treated or not treated with Ru486 (representative images, left) and the locations of GFP⁺ cells were recorded in Ru486-treated mice (right). Scale bar, 50 μm. b, Representative immunostaining for GFP and the suprabasal marker K1 from three mice. Some GFP⁺ cells, which had been HFSCs at the timing of Ru486 treatment, expressed K1 when they moved up to the epidermis. Anagen induction was performed by hair depilation using hair plucking. Scale bar, 50 μm (left), 30 μm (bottom right). Top right, timing of experiment. c, Representative whole-mount staining for K14 and K1 of hair follicles from three mice. K1 expression was higher in anagen hair follicles of HFD-fed mice than in anagen hair follicles of ND-fed mice. Scale bar, 60 μm. d, FACS analysis of dorsal skin; the CD34^highITGA6^highSCA1⁻ population was used as HFSCs for qPCR, microarray analyses and ATAC–sec analysis. e, qPCR analysis for Krt1 in HFSCs from ND- or HFD-fed mice. n = 4 mice, two-tailed unpaired t-test. f, Left, immunostaining for activated CASP3, a marker for apoptosis, and K15 in dorsal skin after hair plucking (pk) or hair removal cream. Right, quantification of hair follicles (HF). n = 3 mice, two-tailed unpaired t-test. g, Whole-mount staining of physiological anagen hair follicles (without any experimental treatment to induce anagen) for K1 and K14. Scale bar, 60 μm. Images are representative of mice aged 25 d (P25; n = 3), 10 m (n = 2) or 24 m (n = 1). Dotted lines show bulge regions. h, Representative cross-section staining of anagen hair follicles from three HFD-fed K15crePR;Rosa-H2B-GFP mice for K1 and GFP. Anagen induction was performed by hair depilation using hair plucking. Scale bar, 50 μm. Arrows in g, h indicate ectopic expression of K1. Data shown as mean ± s.d.

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