Fig 1: Control of GM-CSF Production in Synovial ILCs by IL-2, IL-33, and TLR-9 Ligands(A) Quantity of the active form IL-33 assessed by ELISA (Biolegend, Mouse IL-33 ELISA kit) in indicated tissue homogenates.(B) Quantitative RT-PCR for Il33 expression and ELISA for IL-33 protein in arthritic or control joints. Symbols represent individual mice.(C) Production of cytokines by synovial ILCs. Synovial ILCs (5 × 103) from arthritic joints were purified and cultured for 24 hr with rhIL-2 (20 U/mL), rmIL-7 (20 ng/mL), and rmIL-33 (20 ng/mL) alone or in combination. The concentration of IL-5, IL-13, and GM-CSF in the supernatant was measured (n = 3).(D) Quantitative RT-PCR analysis of the expression of indicated TLR genes in naive CD4+ T cells and synovial ILCs as shown in Figure 4M (n = 3).(E) GM-CSF production by synovial ILCs. Synovial ILCs (5 × 103) were cultured for 24 hr with poly(I:C) (1 µg/mL), LPS (1 µg/mL), and CpG DNA (1 µM) alone or in combination with rmIL-33 (20 ng/mL). The concentration of GM-CSF in the supernatant was measured (n = 3).**p < 0.01. Data are representative of at least two independent experiments. Horizontal bars indicate the means in (A) and (B). Vertical bars indicate SD in (C)–(E).
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