Journal: Experimental Dermatology

Article Title: Interferon‐γ downregulates tight junction function, which is rescued by interleukin‐17A

doi: 10.1111/exd.14425

Figure Lengend Snippet: Dose‐ and time‐dependent inhibition of CLDN1 expression by IFN‐γ in normal human epidermal keratinocytes (NHEKs). (A) Effects of IL‐4, IL‐17A, IFN‐γ and TNF‐α (30 ng ml −1 ) on CLDN1 expression in NHEK cultures incubated with the cytokines in a 24‐well plate for 48 h. Samples were prepared from another three wells ( n = 3). (B) Dose‐dependent inhibition of CLDN1 expression by IFN‐γ (3–100 ng ml −1 ) in NHEKs cultured with or without IFN‐γ for 48 h ( n = 2). (C) Time‐dependent inhibition of CLDN1 expression in NHEKs treated with 30 ng ml −1 IFN‐γ for 0–48 h ( n = 1). Western blotting performed with anti‐CLDN1 and anti‐GAPDH antibodies. Examples of typical blots in duplicate are illustrated. The expression level of CLDN1 was quantified by densitometry and normalized to GAPDH; the ratio is shown directly under each blot. Non‐treated control (0) is 1.00. The statistical analysis is shown in Figure . The results are representative of more than three independent experiments. (D) Changes in the relative CLDN1 mRNA expression in NHEKs cultured in a 24‐well plate with or without IFN‐γ (30 ng ml −1 ) for 3 or 6 h. (E) Changes in the relative CLDN1 mRNA expression in NHEKs treated with 2–50 ng ml −1 IFN‐γ for 6 h. mRNA expression was measured using qRT‐PCR. Data are presented as the mean ± SD ( n = 4). Data were analysed using the Dunnett's test (* p < 0.05; ** p < 0.01 vs. control (0 ng ml −1 ))

Article Snippet: Human recombinant cytokines (IL‐4, IL‐17A, IFN‐γ and TNF‐α) were purchased from FUJIFILM Wako Pure Chemical (Osaka, Japan).

Techniques: Inhibition, Expressing, Incubation, Cell Culture, Western Blot, Control, Quantitative RT-PCR