Fig 2: Dectin-1 induces avß8 expression via IFN-ß, which is critical for TGF-ß activation and non-pathogenic TH17 polarization.a,b, SEAP assay on supernatant of HEK-Blue TGF-ß reporter cells for quantification of active TGF-ß in the supernatant of unstimulated DCs or after stimulation of DCs with curdlan or C. albicans, in the presence of blocking avß1, avß3, avß5, avß6 and avß8 antibodies or isotype (IgG1 and IgG2a) control antibodies (a) or after transduction with non-targeting (control) or specific siRNAs to silence av (ITGAV) or ß8 (ITGB8) expression (b) at 24 h (a, IgG1, avß1, avß3, avß5, avß6 Ab n = 2, IgG2a, avß8 Ab n = 5; b, n = 4). Data in a,b represent mean ± s.d. of independent donors. **P < 0.01, *P < 0.05 (paired, two-tailed Student’s t-test), calculated between untreated and treated (a) or control and specific siRNA-transduced (b) samples that were likewise stimulated. c,d,g,h, Flow cytometry analyses by staining for av (c) or ß8 (d,g,h) expression (FI) in unstimulated DCs or after stimulation of DCs with curdlan or C. albicans (d), in the presence of blocking dectin-1 or IFN-a/ßR antibodies (g) or after silencing of IRF1 or IRF5 expression (h), at indicated times (c, n = 2; d,g, n = 3; h, n = 4). Isotype indicates negative control staining. Representative histograms for independent donors are shown. e,f, Real-time PCR analyses of ITGB8 relative mRNA levels in unstimulated DCs or after stimulation of DCs with curdlan or C. albicans (e), in the presence of blocking dectin-1 or IFN-a/ßR antibodies (f) or after silencing of IRF1 or IRF5 expression (f), at indicated times (e, curdlan n = 8, C. albicans n = 4) or 6 h (f, n = 4). Results from real-time PCR were normalized to the expression of reference household gene GAPDH and shown relative to 6 h curdlan stimulation. Data in e,f represent mean ± s.d. of independent donors. **P < 0.01 (paired, two-tailed Student’s t-test), calculated between untreated and treated or control and specific siRNA-transduced (f) samples that were likewise stimulated. i, Real-time PCR analyses of normalized mRNA levels for indicated genes in IL-17+ T cells, isolated by flow cytometry-based sorting after TH polarization by co-culture of memory CD4+ T cells with DCs primed for 48 h with curdlan or C. albicans, in the presence of blocking avß8 antibodies (during DC stimulation) and/or rhTGF-ß1 (during DC-T co-culture), at day 11–14 (n = 3). Results from real-time PCR were quantified using standard curves for all genes and normalized to the expression of reference household gene ACTB. Data represent mean (number within square) of independent donors: more detailed data are presented in Extended Data Fig. 6. Color legends are as described in Fig. 2.Source data