Figure 4: Epidermal loss of PLCδ1 results in IL-17 upregulation and granulocytosis.

(a) Genomic structure of the PLCδ1 gene (WT). Exons are indicated by filled boxes. The structures of the targeting vector (TV) for disrupting the mouse PLCδ1, targeted allele (MT), allele after FLP recombination (After FLP), and allele after Cre recombination (After Cre) are displayed. (b) PCR genotyping of control and cKO mice. (c) Immunoblotting of PLCδ1 and β-actin in tissues from control, PLCδ1^−/− (KO), and cKO mice. (d) IL-17 mRNA expression in skin. KO ILN was used as positive control for IL-17 expression. Results are displayed as arbitrary units (expression in KO ILN=1). Mean±s.e.m. (n=3). (e) IL-17 mRNA expression in epidermis (epi) and whole skin (skin). Results are displayed as arbitrary units (expression in cKO whole skin=1). Mean±s.e.m. (n=3). (f) Representative FACS profiles of IL-17 and CD3 of epidermis (n=3). Three independent experiments were performed. (g) Macroscopic appearance of ILNs. Scale bar=2 mm. (h) Absolute numbers of IL-17⁺ cells in ILNs. Mean±s.e.m. (n=6). (i) Representative FACS profiles of γδ-TCR and intracellular IL-17 in ILNs (n=6). Cells were stimulated. Five independent experiments were performed. (j) IL-17 mRNA expression in the ILNs. Results are listed as arbitrary units (expression in control mice=1). Mean±s.e.m. (n=3). (k) G-CSF expression in Swiss 3T3 cells treated with skin-draining lymph-node CM in the presence of anti-IL-17 neutralizing antibody or isotype control. Results are displayed as arbitrary units (expression in cells treated with control skin-draining lymph-node CM and isotype control=1). Mean±s.e.m. (n=4). (l) IL-17 concentration in serum. Mean±s.e.m. (n=10). (m) G-CSF concentration in the serum. Mean±s.e.m. (n=6). (n) Representative FACS profiles of CD11b⁺ Gr-1⁺ granulocytes (n=8). (d, e, j, k) All values are normalized to GAPDH. (d–n) 8–12-week-old mice were used. Statistical significance was assessed using a Student′s t-test. *P<0.05; **P<0.01. ND, not detected.