Extended Data Fig. 2: NFI-dKO Mice Exhibit Features of Primary Cicatricial Alopecia.
a, Immunofluorescence showing HF degeneration in NFI-dKO mice at 2 months post-TAM. YFP labels Sox9-CreER-targeted HFs. K6 labels inner bulge cells anchoring the hair. b, Immunofluorescence showing hyperkeratosis (K10) and follicular plugging of infundibulum in NFI-dKO HFs. K5 marks basal epithelial cells. c, Loss of PPARG, SCD1 and ADIPOPHILIN, lipid-related markers of mature sebocytes, in NFI-dKO follicles. Mean and standard deviation are shown. 30 HFs per genotype, pooled from n = 3 mice. P value is from unpaired, two-tailed t-test. d, Hematoxylin & Eosin image of NFI-dKO skin at 2 months post-TAM. Note follicular remnants and fibrous tracts (arrows). e, Immunofluorescence and quantifications of active CASPASE3+ (apoptotic) cells in NFI-dKO HFs. Mean and standard deviation are shown. For 2 weeks data, n = 80 HFs per genotype (total, pooled from 4 mice). For 2 months data, n = 59 HFs (WT) or n = 74 HFs (NFI-dKO), (total, pooled from 3 mice). P values are from unpaired, two-tailed t-test. f, (left) Flow cytometry analysis of immune cells at 2 months post-NFI deletion. Mean and standard deviation are shown. n = 4 mice/genotype. P values are from unpaired, two-tailed t-test. (right) Immunofluorescence analysis of FOXP3+ regulatory T-cells (Tregs) around the HF bulge niche. g, Fluorescence in situ hybridization (FISH) of pan-bacterial 16S rRNA (rainbow colors) in cleared skin whole-mounts, co-labeled for DAPI (gray) at 2 months post-TAM. Spot analysis of 16S-FISH signal was used to quantify bacterial load per μm3 of skin. Mean and standard deviation are shown. n = 10 (WT) and 16 (NFI-dKO) HFs from 2 biologically independent mice/genotype. Scale bars = 20 μm, unless otherwise specified. Bu, bulge. Inf, Infundibulum. Dashed lines, HF-dermal border. For a-d and f, at least three biological replicates were used; representative images are shown. See also Source Data.
