Figure 4.

Rat CLEC-1–deficient BMDCs enhance Th17-cell activation. (A) BMDCs from WT and CLEC-1–deficient rats were stimulated with TLR 4-L or zymosan for 24 hours, and CD80, CD86, and Class I and II major histocompatibility complex (MHC) were assessed by flow cytometry. Data were expressed in histograms as mean ± SEM of 6 independent experiments. (B) BMDCs were stimulated with TLR4-L or zymosan for 8 hours, and Il12p40, Il12p35, Il23p19, Il6, Il10, and Tgfb1 were assessed by qRT-PCR. Results were expressed in histograms as mean ± SEM of 6 independent experiments and were expressed in AU of specific cytokine/Hprt ratio. (C) BMDCs were incubated for 4 days in MLR with allogeneic purified CD4⁺ T cells. (i) Histogram and representative staining of proliferation (CFSE dilution) assessed in CD4⁺ T cells by flow cytometry, and (ii) histogram and representative dot plots of percentage of IL-17⁺ and IFN-γ⁺ cells among gated CD4⁺ T cells assessed by flow cytometry. Data were expressed in histograms as mean ± SEM of 6 independent experiments. *P < .05; **P < .01. KO, knockout; MFI, mean fluorescence intensity; mRNA, messenger RNA; NS, nonstimulated.