Supplementary Figure 1: IL-6 reporter mice.
From: Trans-presentation of IL-6 by dendritic cells is required for the priming of pathogenic TH17 cells
(a) Targeted Il6 locus. The reporter cassette including a floxed stop cassette was introduced into exon 2 of the Il6 locus. Since the locus is disrupted by this knock-in construct, IL-6 reporter mice were bred heterozygously and compared with Il6+/- mice since Il6 produces a gene dose effect (data not shown). (b) Bone marrow derived dendritic cells (BMDCs) were prepared from CMV-Cre x Il6RD/wt mice and stimulated in vitro with CpG. Cerulean was expressed in the cytoplasm (left) and Thy1.1 at the cell surface (middle, merge right) as expected. Confocal microphotographs, Scale bar 10 μm. (c) Il6RD/wt control BMDCs or BMDCs prepared from IL-6 reporter mice (CMV- Cre x Il6RD/wt) were stimulated with LPS followed by flow cytometric assessment of Thy1.1 expression. (d-f) Correlation of Thy1.1 expression and IL-6 expression in BMDCs. BMDCs were prepared from different mouse strains as indicated and stimulated with LPS followed by analysis of Thy1.1 expression (d) and IL-6 production as measured by ELISA (e). (f) Co-expression of IL-6 and Thy1.1. Control BMDCs (Il6RD/wt) and IL-6 reporter BMDCs (CD11c-Cre x Il6RD/wt) were stimulated with CpG for 6 h in the presence of Brefeldin A for the last 2 h followed by combined surface staining for Thy1.1 and intracellular staining for IL-6.
