Fig. 2
IFNγ promotes antigen cross-presentation in OPCs. Isolated and PDGF (20 ng/mL) expanded postnatal rat OPCs (P4-P6) were differentiated with T3 (white), T3+IFNγ (10 ng/mL; charcoal), T3+IL17 (50 ng/mL; blue), or T3+IFNγ+IL-17 (black) for 96 h prior to assessment using qPCR (a) and ICC staining for MBP and Olig2 (b). Isolated and PDGF (20 ng/mL) expanded postnatal rat OPCs (P4-P6) were left undifferentiated (0 h; white) or differentiated with T3 (10 nM), IFNγ (10 ng/mL), T3 + IFNγ, for 8 hrs (light gray), 24 hrs (medium gray), 48 h (charcoal) and 96 hrs (black) to compare gene expression between groups while obtaining information at multiple time points. c–e Affymetrix gene arrays analysis was performed from three independent biological replicates. c Venn diagram summarizing the number and overlap of up-regulated (left) and down-regulated (right) genes based on time point. d Global heat map of all probes clustered into three gene expression patterns comparing PDGF, T3, and T3+IFNγ at all time points. e Targeted heat map comparing PDGF, T3, IFNγ, and T3+IFNγ at all time points. Displayed genes were identified from GSEA analysis (supplement 3). f Quantitative PCR validation of OPCs differentiated with T3 (white), T3+IFNγ (10 ng/mL; gray), or T3+IL17 (50 ng/mL; blue) for 96 h prior to assessment. Error bars represent the standard deviation from three independent primary isolations and experiments. Significance for qPCR analysis was determined by one-way ANOVA analysis followed by Dunnett’s multiple comparison analysis where T3+IFNγ or T3+IL17 were compared to T3 alone control (P*≤0.05, **≤0.01, ***≤0.001, ****≤0.0001)
