rabbit anti human il 22r antibody  (R&D Systems)

Journal: Oncotarget

Article Title: Interleukin-22 participates in the inflammatory process of vitiligo

doi: 10.18632/oncotarget.22644

Figure Lengend Snippet: (A) To block IL-22R-mediated signaling, pre-treatment of HaCaT cells with 50 μg/ml of anti-IL-22R blocking antibody for 2 h prior to IL-22 stimulation resulted in reduced NLRP3 and caspase-1 expression (B) as well as a consequent reduction in IL-1β secretion as measured by ELISA. (C) To reduce NLRP3 expression in HaCaT cells, we transfected NLRP3 siRNA or control siRNA into HaCaT cells. The level of protein was quantified to evaluate the knockdown efficiency of the endogenous NLRP3 protein in HaCaT cells. Data from three independent experiments for protein expression are shown. (D) The active form of IL-1β in culture supernatants was quantitated by ELISA. (E) NLRP3 knockdown in HaCaT cells resulted in a reduction in the levels of caspase-1 and the active form of IL-1β in IL-22-treated cells. Representative blots of three independent experiments are shown. Results were normalized against β-actin expression. The data are expressed as means ± SD (i.e. n=3) ( ** P < 0.05). Ab, anti-IL-22R blocking antibody; IgG, isotype antibody for the negative control of IL-22R antibody.

Article Snippet: The cells were pretreated with either rabbit anti-human IL-22R antibody (AF2770; R&D systems) or with the isotype control (AB-108-C; R&D systems) at a concentration of 50 μg/ml for 2 h to block IL-22R in HaCaT cells.

Techniques: Blocking Assay, Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Control, Knockdown, Negative Control

Journal: European journal of immunology

Article Title: IL-17 and IL-22 mediate IL-20 subfamily cytokine production in cultured keratinocytes via increased IL-22 receptor expression.

doi: 10.1002/eji.200939473

Figure Lengend Snippet: Figure 3. IL-20R1, IL-20R2, IL-22R, and IL-10R2 expression in cultured epidermal keratinocytes. (A) Normal epidermis was obtained from nine subjects and cultured keratinocytes were from eight other normal subjects. IL-20R1, IL-20R2, IL-22R, and IL-10R2 expression levels were determined by real-time RT-PCR, with cyclophilin as house keeping control, and compared between normal epidermis (black bar) and cultured keratinocytes (gray bar). Data show mean7SD. po0.05; po0.001. (B) Western blot analysis of IL-20R1, IL-20R2, and IL-22R protein expression in cultured keratinocytes. (C) IL-22R expression levels were analyzed by flow cytometry (isotype control Ab: shaded area; anti-IL-22R Ab: thin line). Infection of AxIL22R at an MOI of 5 for 48 h increased IL-22R expression in keratinocytes (thick line), but not AxLacZ (dotted line). Data are representative of two independent experiments. (D) Uninfected control cultures or keratinocytes infected with AxIL22R were treated with 10 ng/mL IL-19, IL-20, IL-24, and IL-22. STAT-3 phosphorylation at 15 min or 24 h after stimulation was determined by Western blot. Data are representative of three independent experiments.

Article Snippet: The following primary antibodies were used: goat anti-IL-19 antibody (AF1035, R&D Systems), mouse anti-IL-20 antibody (Abcam, Tokyo, Japan), mouse anti-IL-24 antibody (MAB 19651, R&D Systems), and goat anti-IL-22R antibody (AF2770, R&D Systems).

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Control, Western Blot, Cytometry, Infection, Phospho-proteomics

Journal: European journal of immunology

Article Title: IL-17 and IL-22 mediate IL-20 subfamily cytokine production in cultured keratinocytes via increased IL-22 receptor expression.

doi: 10.1002/eji.200939473

Figure Lengend Snippet: Figure 6. IL-22R expression in the epidermis of normal and psoriatic skin. (A) Total RNA was extracted from the epidermis of nine normal and 11 psoriatic skin samples. IL-20R1, IL-20R2, IL-22R, and IL-10R2 mRNA expression levels were determined by real-time RT-PCR, with cyclophilin as house keeping control. Results were expressed as relative expression levels of mRNA (gray bar) compared with that in normal epidermis (black bar). Data show mean7SD.po0.05. (B) IL-22R immunostaining of four normal and six psoriatic skin samples. In four psoriatic skin samples, fluorescence intensity increased markedly as a result of IL-22R over-expression. Scale bar, 50 mm.

Article Snippet: The following primary antibodies were used: goat anti-IL-19 antibody (AF1035, R&D Systems), mouse anti-IL-20 antibody (Abcam, Tokyo, Japan), mouse anti-IL-24 antibody (MAB 19651, R&D Systems), and goat anti-IL-22R antibody (AF2770, R&D Systems).

Techniques: Expressing, Quantitative RT-PCR, Control, Immunostaining, Over Expression

Journal: European journal of immunology

Article Title: IL-17 and IL-22 mediate IL-20 subfamily cytokine production in cultured keratinocytes via increased IL-22 receptor expression.

doi: 10.1002/eji.200939473

Figure Lengend Snippet: Figure 7. Effect of IL-20, IL-24, and IL-22 on cytokine production. (A) Cultured keratinocytes transduced with (black bar) or without (gray bar) IL-22R by an adenovirus vector were treated for 36 h with IL-20 (10 ng/mL), IL-24 (10 ng/mL), or IL-22 (10 ng/mL) in combination with or without IL-17 (10 ng/mL). The levels of mRNA expression were corrected using cyclophilin expression relative to the untreated cells (control) level. (B) Cultured keratinocytes transduced with IL-22R by an adenovirus vector were treated with IL-20 (10 ng/mL), IL-24 (10 ng/mL), or IL-22 (10 ng/mL) for 48 h in combination with (10 ng/mL) or without IL-17. Culture media were collected, and MIP-3a and HB-EGF were measured by ELISA. The analysis was performed in triplicate. Data show mean7SD. Data are representative of three independent experiments. po0.05; po0.01.

Article Snippet: The following primary antibodies were used: goat anti-IL-19 antibody (AF1035, R&D Systems), mouse anti-IL-20 antibody (Abcam, Tokyo, Japan), mouse anti-IL-24 antibody (MAB 19651, R&D Systems), and goat anti-IL-22R antibody (AF2770, R&D Systems).

Techniques: Cell Culture, Transduction, Plasmid Preparation, Expressing, Control, Enzyme-linked Immunosorbent Assay

Journal: European journal of immunology

Article Title: IL-17 and IL-22 mediate IL-20 subfamily cytokine production in cultured keratinocytes via increased IL-22 receptor expression.

doi: 10.1002/eji.200939473

Figure Lengend Snippet: Figure 8. Role of IL-22R and the IL-20 subfamily in psoriasis. In psoriatic skin, IL-22R expression in the epidermis is elevated compared with normal skin. IL-22 and IL-17 produced by Th17 lymphocytes induce abundant expression of the IL-20 subfamily and chemokines such as MIP-3a and IL-8, and HB-EGF compared with normal skin. MIP- 3a and IL-8 recruit further Th17 lymphocytes and neutrophils, respectively. HB-EGF may induce epidermal hyperproliferation. Surrounding keratinocytes treated with IL-20 and IL-24 strongly responded to IL-17 and produced more MIP-3a, IL-8, and HB-EGF. Phosphorylation of STAT3 was observed in the nuclei of keratinocytes stimulated with IL-22 or IL-20 subfamily members.

Article Snippet: The following primary antibodies were used: goat anti-IL-19 antibody (AF1035, R&D Systems), mouse anti-IL-20 antibody (Abcam, Tokyo, Japan), mouse anti-IL-24 antibody (MAB 19651, R&D Systems), and goat anti-IL-22R antibody (AF2770, R&D Systems).

Techniques: Expressing, Produced, Phospho-proteomics