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. 2021 Jan 29;7(5):eabe0337. doi: 10.1126/sciadv.abe0337

Fig. 1. Corticosteroidogenic key enzymes are expressed in human and mouse skin.

Fig. 1

(A) Immunohistochemistry with anti-human CYP11B1 or rabbit immunoglobulin G (IgG) of human skin from healthy donors or patients with lesional AD or psoriasis. Representative images from healthy donors (n = 5), patients with lesional AD (n = 8), or lesional psoriasis (n = 10). Scale bar, 100 μm. (B) Immunofluorescence for CYP11A1 (green) and CD45 (red) or rabbit and rat isotype IgG on frozen human skin sections from healthy donors or patients with lesional AD or psoriasis. Cell nuclei are stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Representative images from three individual donors and patients are shown. White dashed line indicates dermal-epidermal junction. Scale bar, 50 μm. (C) Reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis of CYP11B1, HSD11B1, and CYP11A1 in laser capture microdissected epidermis from frozen human skin sections of healthy donors (HD), patients with lesional AD (AD) or psoriasis (PS). Expression was normalized to GAPDH, and data are depicted as 2(−∆Ct). Columns represent means ± SEM (n = 2 to 3 individuals per group) of one experiment. (D) Corticosterone radioimmunoassay from untreated (UT) or metyrapone (MET)–treated ex vivo mouse tissue culture in response to phosphate-buffered saline (PBS) or LPS. Symbols represent individual animals. Columns show means ± SEM (n = 4 to 6 per group), pooled from two independent experiments. (E) RNA expression in immortalized C57BL/6 keratinocytes that were untreated or treated with 1 μM ACTH or 20 μM forskolin (FSK) for overnight. Expression was normalized to Actb, and data are depicted as fold change to untreated samples. Data represent means ± SEM (n = 4 to 8 per group), pooled from two to three independent experiments. Statistical significance for (E) was determined using the Kruskal-Wallis test with Dunn’s multiple comparisons test and the ordinary one-way analysis of variance (ANOVA) with Dunn’s multiple comparisons test for Hsd11b1 expression analysis.