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. 2017 Jun 28;8:722. doi: 10.3389/fimmu.2017.00722

Figure 3.

Figure 3

Overexpression of the IL-33 gene in cultured keratinocytes and the skin selective hK14mIL33tg mouse. (A) Schematic representation of the transgene DNA. An active full-length form of mouse IL-33 cDNA was placed downstream of the human keratin 14 promoter. The transgene construct also contained a rabbit β-globin intron sequence and a keratin 14 polyadenylation signal for the stable processing of the transcripts. (B) Transfection of the human keratinocyte cell line HaCaT with mock (M) or hK14mIL-33 (Tg). After 48 h or 72 h, supernatant (S) and cell pellets were stained for IL-33. (C) Immunhistochemistry of hK14mIL-33 or mock transfected HaCaTs. Nuclear staining in the lower panel (DAB, brown dye 40×). (D) No cutaneous manifestations of hK14mIL33tg mice up to 6 months. (E) Immunhistochemistry of IL-33 in the epidermis of wild-type (WT) and hK14mIL33tg (Tg) back skins. (F) The skin-selective expression of the IL-33 gene in hK14mIL33tg mice. Total RNAs from various organs of mice were used as templates for quantitative real-time PCR. Each bar shows the expression of the IL-33 gene in hK14mIL33tg mice relative to WT mice, data represent mean and SEM of n = 3 (except liver and stomach; n = 1). Other data are representative of three independent experiments.