Figure 6: cKO skin exhibits features of human inflammatory skin disease.

(a) Immunoblotting of PLCδ1 and β-actin in whole skin or epidermis from non-treated (non) or IMQ-treated (IMQ) mice. (b) HE stained dorsal skin sections. Scale bar, 50 μm. The control skin has a normal epidermis, whereas the cKO skin has a thickened epidermis. (c) The skin was stained with antibodies against CD3 (green) or F4/80 (green) and Hoechst (blue). Scale bar,100 μm. (d) CCR6, CCL20, β-defensin3 (BD3), TNF α, IL-6, and S100A9 mRNA expression in skin determined by real-time RT–PCR. All values are normalized to GAPDH. Results are displayed as arbitrary units (expression in skin of control mice=1). Mean±s.e.m. (n=5). (e) Immunoblotting for total and phosphorylated STAT3 (pSTAT3) in skin. β-actin was included as a loading control. (f) Skin stained with antibody against phosphorylated STAT3. Dotted lines denote dermal–epidermal border. Scale bar, 20 μm. Nuclear staining of phosphorylated STAT3 is indicated by arrows. (g) VEGF mRNA expression in skin determined by real-time RT–PCR. All values are normalized to GAPDH. Results are displayed as arbitrary units (expression in skin of control mice=1). Mean±s.e.m. (n=5). (h) Skin stained with the antibody against CD31 (red) and Hoechst (blue). Hair shafts show nonspecific auto-fluorescence (red). Scale bar, 100 μm. (i) Skin from four non-psoriatic volunteers and four patients with psoriasis were stained with antibody against human PLCδ1 (brown). Dotted lines denote dermal–epidermal border. Scale bar, 100 μm. Body sites of each skin samples were as follows: control; arm, waist, back, and back (indicated from the top panel to the bottom panel). Psoriasis; arm, abdominal, leg, and leg (indicated from the top panel to the bottom panel). (a, c–h) 8–12-week-old mice were used. (b–h) Untreated IMQs were used. The data presented in (b, c, e, f, h) are representative of analyses of three mice per genotype. The data presented in (a) is representative of analyses of two mice per genotype. Statistical significance was assessed using a Student′s t-test. *P<0.05; **P<0.01.