The elevation of GATA-3/T-bet ratio in EAE-afflicted wildtype mice by 1,25(OH)2D3 did not occur in STAT6 null mice. A) At sacrifice of the mice in Figure 4, spleens were removed for RNA whereby qPCR was performed. The GATA-3/T-bet ratios were increased by 1,25(OH)2D3 (denoted as Vit D) in wildtype (WT) mice but this elevation did not occur in STAT6 null (knockout, KO) mice. N = 5 per group, mean ± SEM, repeated twice. For panels B and C, spleen and lymph nodes from the animals sacrificed in Figure 4 were processed by Ficoll gradient centrifugation for lymphocytes and these were left untreated, or were treated ex vivo for 3 days with MOG (peptide 35-55), 1,25(OH)2D3 (denoted as Vit D), or MOG plus 1,25(OH)2D3. In wildtype mice (panel B) that were treated in vivo with vehicle or 1,25(OH)2D3, the ex vivo exposure to 1,25(OH)2D3, or MOG plus 1,25(OH)2D3, increased the GATA-3/T-bet ratio; importantly, this increase did not occur in STAT6 KO mice (panel C) whether they were exposed in vivo to vehicle or 1,25(OH)2D3. Bars are mean ± SEM, N = 5 per group, repeated twice.