1. Foxc1 Ablated Mice Are Anhidrotic and Recapitulate Features of Human Miliaria Sweat Retention Disorder 
Chang-Yi Cui, Ryuga Ishii, Dean P. Campbell, Marc Michel, Yulan Piao, Tsutomu Kume, David Schlessinger 
Journal of Investigative Dermatology 
Volume 137, Issue 1, Pages (January 2017)
DOI: /j.jid
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2. Figure 1
Skin-specific Foxc1 cKO mice are hypohidrotic. (a) Iodine-starch sweat test. Black dots represent sweating spots. Foxc1 cKO mice show severe hypohidrosis compared with wild-type (WT) littermate. (b) Localization of Foxc1 in sweat glands. Foxc1-positive cells (red) are located exclusively in nuclei of luminal duct cells (broken circles), but not in basal duct cells (arrows) or secretory portion. Foxc1 cKO mice are devoid of signal. Blue, DAPI staining. Scale bar = 15 μm.
Journal of Investigative Dermatology  , 38-45DOI: ( /j.jid )
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3. Figure 2
Morphologic analyses of Foxc1 cKO sweat glands. (a) Epidermal sweat ducts were blocked by parakeratotic plugs (upper-right panel; cell nuclei can be spotted in the plugs) or hyperkeratotic plugs (lower-right panel). Small panels at right show higher magnifications. Epidermal sweat ducts were empty in wild-type (WT) controls (left panels). (b) Dermal sweat ducts were blocked by hyperkeratotic plugs in upper (upper-right panel) and lower (lower-right panel) portions. Note: compared with WT controls (left panels), dermal ducts in Foxc1 cKO mice were significantly dilated (surrounded by arrows). (c) Lumens in secretory portion were also significantly dilated in Foxc1 cKO mice (upper-right panel, compared with upper-left). Further, secretory cells surrounding dilated secretory lumens were flattened in the cKO mice (arrows in the lower-right panel, compared with the lower-left panel). (d) Illustration of a sweat gland. Sweat glands consist of a short helical epidermal duct, a relatively straight dermal duct, and a coiled secretory portion. Sweat duct blockage in Foxc1 cKO mice was found in the epidermal duct in stratum corneum, lower epidermis, and deeper dermal duct (red dots). (e) Small blisters (upper-right panel) and papules (lower-right panel) were formed in digit tips of Foxc1 cKO mice, but not in WT controls. Scale bars = 50 μm.
Journal of Investigative Dermatology  , 38-45DOI: ( /j.jid )
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4. Figure 3
Ectopic expression of differentiation markers in sweat duct luminal cells in the Foxc1 cKO mice. (a) Krt8 (red) was highly expressed in luminal cells of sweat ducts in Foxc1 cKO mice, but not in wild-type (WT) controls. Krt14 (green) was expressed in basal duct cells in both WT and Foxc1 cKO mice. (b) Sprr2 (a+b) (red) were intensely expressed in luminal duct cells in Foxc1 cKO mice (arrows), but were absent in WT sweat ducts. (c) Loricrin (green) was strongly expressed in the granular and upper spinal layers of skin epidermis, but absent in sweat glands of both WT and Foxc1 cKO mice. Blue, DAPI staining. Scale bars = 20 μm.
Journal of Investigative Dermatology  , 38-45DOI: ( /j.jid )
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5. Figure 4
Expression profiling revealed the upregulation of a set of terminal differentiation markers in Foxc1 cKO sweat glands. (a) A scatter plot shows upregulated or downregulated genes in Foxc1 cKO sweat glands. A total of 185 genes were significantly affected in Foxc1 cKO mice (157 genes up [red] and 28 genes down [purple]). In Foxc1 cKO mice, terminal differentiation markers, including Sprr2a, Sprr2b, Sprr2f, S100a8, S100a9, Krt42, and Krtap13-1, were significantly upregulated, and Foxc1 was sharply downregulated. (b) Functional annotation revealed the upregulation of terminal differentiation markers and immune responsive genes in Foxc1 cKO mice. (c) Quantitative real-time reverse transcriptase-PCR assays confirmed significant upregulation of terminal differentiation markers in Foxc1 cKO sweat glands.
Journal of Investigative Dermatology  , 38-45DOI: ( /j.jid )
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6. Figure 5
Foxc1 represses Sprr2a expression in keratinocytes. (a) Immunostaining shows that the 308 mouse keratinocytes highly express Sprr2a (green). Cells transfected with Foxc1 (red) lack Sprr2a expression. Scale bar = 25 μm. Right panel: almost all counted Foxc1-positive cells are Sprr2a negative (F+/S–). (b) Left panel: Sprr2a constructs for luciferase assay. Right panel: Foxc1 represses promoter activity of Sprr2a, and the 0.5–0.3 kb fragment is responsible for the repression. (c) Left panel: sequence of the 0.5–0.3 kb fragment. Red indicates a partial Fox-binding site. Right panel: chromatin immunoprecipitation assay shows that Foxc1 binds to the 0.5–0.3 kb fragment. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; TG, transgene; TSS, transcription start site.
Journal of Investigative Dermatology  , 38-45DOI: ( /j.jid )
Copyright © 2016 The Authors Terms and Conditions

7. Figure 6
A schematic representation of Foxc1 action in sweat glands. Foxc1 is expressed in sweat duct luminal cells, and its ablation causes ectopic expression of terminal differentiation markers including Sprr proteins, S100a proteins, and several keratin and keratin-associated proteins, resulting in hyperkeratotic or parakeratotic plug formation in sweat duct. A miliaria-like phenotype is caused by sweat duct blockage by keratotic plugs.
Journal of Investigative Dermatology  , 38-45DOI: ( /j.jid )
Copyright © 2016 The Authors Terms and Conditions