Figure 8. AM580-induced differentiation of MDDC enhances IL-22BP expression.

(A) MDDC were differentiated in the absence or presence of AM580 added at day 0 or day 4 of culture. IL-22BP gene expression was analyzed by RT-qPCR. Bars represent mean ± SEM of fold change compared to GM-CSF/IL-4 derived MDDC, of IL-22BP gene to HPRT, expression as determined by the 2^−ΔΔCt method of relative quantification (n=4). (B) Cells were harvested at the indicated times during MDDC differentiation in the presence or the absence of AM580 added at day 0. IL-22BP gene expression was analyzed by RT-qPCR. Each point represents mean ± SEM of fold change compared to GM-CSF/IL-4 derived MDDC, of IL-22BP gene to HPRT expression as determined by the 2^−ΔΔCt method of relative quantification (n=4). (C) MDDC and AM580-diferentiated MDCC were stained for IL-22BP as described in figure 6. (D) MDDC were differentiated in the absence or presence of AM580 added at day 0 or day 4 of culture. Cell surface markers were analyzed by flow cytometry. Red histograms, isotype control staining; blue histrograms, antibody staining. Data are representative of at least four independent experiments. (E) MDDC were differentiated in the absence or presence of RA and specific inhibitors of RA nuclear receptors added at d0. IL-22BP gene expression was analyzed by RT-qPCR. Bars represent mean ± SEM of fold change compared to GM-CSF/IL-4 derived MDDC, of IL-22BP gene to HPRT, expression as determined by the 2^−ΔΔCt method of relative quantification (n=4). (F) MDDC were differentiated in the presence or not of retinal and/or DEAB, a selective inhibitor of RALDH2. IL-22BP gene expression was analyzed by RT-qPCR. Bars represent mean ± SEM of fold change compared to GM-CSF/IL-4 derived MDDC, of IL-22BP gene to HPRT, expression as determined by the 2^−ΔΔCt method of relative quantification (n=3). (G) RALDH2 gene expression was analyzed by RT-qPCR in rat CD172⁺ CD4⁺ spDCs and CD172^high ilDCs. Bars represent mean ± SEM of IL-22BP gene to HPRT, expression as determined by the 2^−ΔΔCt method of relative quantification (n=3). * p<0.05.