Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Identification of functional roles for both IL-17RB and IL-17RA in mediating IL-25-induced activities.

doi: 10.4049/jimmunol.181.6.4299

Figure Lengend Snippet: FIGURE 4. Intranasal IL-25 administration did not induce histological signs of lung inflammation in IL-17RB KO or IL-17RA KO mice. WT mice (A and B), IL-17RB KO mice (C and D), and IL-17RA KO mice (E and F) (n 5 per group) were i.n. dosed with vehicle for 4 days, and lungs were harvested for histological analysis 24 h after the final dose. WT mice (G and H), IL-17RB KO mice (I and J), and IL-17RA KO mice (K and L) (n 5 per group) were i.n. dosed with 0.5 g of IL-25 for 4 days, and lungs were harvested for histological analysis 24 h after the final dose. Formalin-fixed lung tissue sections were stained with H&E (A–L) or PAS (M and N) for analysis. Goblet cell hyperplasia was assessed in PAS-stained sections as described in Materials and Methods. Original magnification, 20 in A, C, E, G, I, and K. Original magnification, 400 in B, D, F, H, J, and L. Statistical analyses of WT and KO mice were performed using a nonparametric one-way ANOVA.

Article Snippet: The goat anti-human IL-17RA Ab (R&D Systems; catalog no. AF177) and mouse anti-human IL-17A mAb clone 41809 (R&D Systems; catalog no. MAB317) are reported by the manufacturer to inhibit IL-17A-induced IL-6 production by normal human dermal fibroblasts.

Techniques: Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Identification of functional roles for both IL-17RB and IL-17RA in mediating IL-25-induced activities.

doi: 10.4049/jimmunol.181.6.4299

Figure Lengend Snippet: FIGURE 6. Abs to IL-17RB, IL-17RA, or IL-25 block IL-25-induced histological signs of pulmonary inflammation. BALB/c mice were i.n. dosed with vehicle (A) or 0.5 g of IL-25 (B–F) for 4 days, and lungs were harvested for histological analysis 24 h after the final dose. Mice (n 5 per group) were treated with control Ab (B) or blocking Abs to IL-17RB (C), IL-25 (D), IL-17RA (E), or IL-17A (F) 3 h before each i.n. dose. Formalin- fixed lung tissue sections were stained with H&E (A–F) or PAS (G) for analysis. A–F, Original magnification, 200. G, Goblet cell hyperplasia was assessed in PAS-stained lung sections as described in Materials and Methods. Statistical analyses were performed using a nonparametric one- way ANOVA.

Article Snippet: The goat anti-human IL-17RA Ab (R&D Systems; catalog no. AF177) and mouse anti-human IL-17A mAb clone 41809 (R&D Systems; catalog no. MAB317) are reported by the manufacturer to inhibit IL-17A-induced IL-6 production by normal human dermal fibroblasts.

Techniques: Blocking Assay, Control, Staining

Journal: International Journal of Molecular Sciences

Article Title: IL-17RA Signaling in Prx1+ Mesenchymal Cells Influences Fracture Healing in Mice

doi: 10.3390/ijms25073751

Figure Lengend Snippet: Conditional deletion of Il17ra in Prx1 + cells leads to subtle changes in bone microarchitecture. ( A ) Prx1-cre ; Il17ra F/F ( Il17ra cKO) were generated to conditionally delete Il17ra in Prx1 + mesenchymal cells by deleting exons 3 and 4 which leads to the early termination of translation. ( B ) Prx1 - cre significantly decreased the Il17ra gene expression in isolated periosteal cells. ( C ) Immunohistochemistry shows less immunoreactivity of IL-17RA within day 14 fracture calluses and the periosteum of Il17ra cKO mice. Scale bar: 50 µm. Representative 3D images of ( D ) femur trabecular bone, ( E ) femur cortical bone, and ( F ) L3 vertebral body trabecular bone. µCT analysis showing ( G ) femur trabecular bone volume fraction (BV/TV), ( H ) femur trabecular thickness (Tb.Th), ( I ) femur cortical thickness (Ct.Th), ( J ) femur cortical porosity (Ct.Po), ( K ) L3 vertebral body trabecular bone volume fraction (BV/TV), and ( L ) L3 vertebral body trabecular thickness (Tb.Th). Abbreviations: PO, periosteum; Prom, promoter; cKO, conditional knockout. Data represent mean ± SD. Student’s unpaired t -test, * p < 0.05, ** p < 0.001.

Article Snippet: Sections were blocked with 10% normal Goat serum for 1 h then incubated overnight with Rabbit Anti-IL-17RA antibody (1:100 in 1.5% normal Goat serum; Abcam, Boston, MA, USA #ab218249) in a humidified chamber placed at 4 °C.

Techniques: Generated, Expressing, Isolation, Immunohistochemistry, Knock-Out

Journal: International Journal of Molecular Sciences

Article Title: IL-17RA Signaling in Prx1+ Mesenchymal Cells Influences Fracture Healing in Mice

doi: 10.3390/ijms25073751

Figure Lengend Snippet: Activation of IL-17RA signaling inhibits osteogenesis. ( A ) IL-17A at 20 and 50 ng/mL inhibited the gene expression of Runx2 , Osx , Cola1 , and Bglap in periosteal cells isolated from control mice. ( B ) IL-17A did not influence expression of Runx2 , Osx , Cola1 , and Bglap in Il 1 7ra cKO periosteal cells. ( C ) Alizarin red-S staining and quantification shows less mineralization in IL-17A-treated control periosteal cells at days 14 and 21 of osteogenic differentiation, but there was no effect on mineralization by Il17ra cKO cells. Dashed line represented a fold-change of 1. Abbreviations: Diff, osteogenic differentiation; Osx , osterix; Cola1 , collagen type 1; Bglap, osteocalcin. Data represent mean ± SD. The two-way ANOVA followed by Tukey’s multiple comparisons, * p < 0.05.

Article Snippet: Sections were blocked with 10% normal Goat serum for 1 h then incubated overnight with Rabbit Anti-IL-17RA antibody (1:100 in 1.5% normal Goat serum; Abcam, Boston, MA, USA #ab218249) in a humidified chamber placed at 4 °C.

Techniques: Activation Assay, Expressing, Isolation, Staining

Journal: International Journal of Molecular Sciences

Article Title: IL-17RA Signaling in Prx1+ Mesenchymal Cells Influences Fracture Healing in Mice

doi: 10.3390/ijms25073751

Figure Lengend Snippet: IL-17RA signaling promotes periosteal progenitor cell migration. Wound healing assay shows that IL-17A (20 ng/mL) promoted cell migration at 12 h. ( Left ): representative images of scratched areas marked by black lines. ( Right ): the semi-quantitative analysis of wound closure was determined by measuring the widths of the wounds. Data represent the mean ± SD. Two-way ANOVA followed by Tukey’s multiple comparisons. Values not sharing a common letter differ significantly, p < 0.05.

Article Snippet: Sections were blocked with 10% normal Goat serum for 1 h then incubated overnight with Rabbit Anti-IL-17RA antibody (1:100 in 1.5% normal Goat serum; Abcam, Boston, MA, USA #ab218249) in a humidified chamber placed at 4 °C.

Techniques: Migration, Wound Healing Assay