Porcn inactivation blocks Wnt signaling and caused alopecia in adult mice, but did not impair hematopoiesis. (A) Inactivation of Porcn downregulated expression of Wnt/β-catenin target gene (Axin2) in skin samples of Rosa-CreER^T2/Porcn^Del mice. Expression was normalized to PGK. The mean values of Porcn^WT samples were set to 1 within each group (n = 4 mice in each group, 2 independent experiments, *P < .05, Mann-Whitney test). (B) Tamoxifen supplementation in chow for 3 weeks caused progressive alopecia in mice. Photo taken 7 weeks after start of tamoxifen. (C) Loss of hair follicles in skin after Porcn inactivation. H&E stain. Scale bar = 200 μm. (D) CBC after deletion of Porcn (5-6 weeks after tamoxifen chow administration). Total WBC and lymphocyte counts did not differ significantly between Rosa-CreER^T2/Porcn^WT (n = 5) and Porcn^Del mice (n = 9) (*P < .05; Mann-Whitney test), whereas neutrophils were modestly increased (representative data of 4 independent experiments, P = .019, Mann-Whitney test). (E) Porcn was downregulated in BM of Porcn-inactivated mice 4 weeks after tamoxifen chow administration. Expression assessed by qRT-PCR was normalized to PGK and β-actin. Individual expression was normalized to mean of respective Porcn^WT samples within the same group (n = 6; 2 independent experiments; *P < .01; Mann-Whitney test). (F) The total number of LT-HSCs (Lin⁻, Sca1⁺, c-Kit⁺, CD150⁺, CD48⁻), ST-HSCs (Lin⁻, Sca1⁺,c-Kit⁺, CD150⁻, CD48⁻), and HPCs (Lin⁻, Sca1⁺,c-Kit⁺, CD150⁻, CD48⁺) did not differ between Rosa-CreER^T2/Porcn^WT and Porcn^Del mice (right panel). Representative flow cytometry data are shown in left panel (n = 6, cell number per 1 leg, 2 independent experiments, P > .05, Mann-Whitney test). (G) Total CLP cells (1 leg) were similar in Rosa-CreER^T2/Porcn^WT and Porcn^Del mice 4 weeks after tamoxifen (n = 6 in each group, P > .05, Mann-Whitney test). (H) Total number of myeloid progenitor cells (1 leg) were similar in Rosa-CreER^T2/Porcn^WT and Porcn^Del mice (middle and right panels) (n = 6 mice, 4 independent experiments, P > .05, Mann-Whitney test). Left panel, The representative gating for CMP, MEP, and GMP. H&E, hematoxylin and eosin; NS, not significant.