FIGURE 3.
Conditioned media from TI stimulated astrocytes suppresses human OPC differentiation. (A) Schematic depicting experiment. (B) Barplot showing Nanoluciferase activity (normalized to media only control) on Y-axis. Each dot represents a well of hiPSC derived astrocytes whose media was placed onto five identical replicate wells of hOPC reporter cells. Columns represent mean of replicate astrocyte samples, dots represent mean of replicate OPC wells, and error bars represent standard error of the mean of replicate OPC wells. (C) Barplot showing nuclear counts following treatment with astrocyte CM, data presented in same format as in (C). (D) Principle component analysis of hOPC reporter RNA libraries comparing samples treated with media only, media only with TI, and CM from NMSC and PwMS derived astrocytes treated with or without TI. (E) Heatmap showing relative expression of genes previously associated with inflammatory OPCs in hOPC reporter cells treated with astrocyte CM (and media only controls). Dotplots showing selected GO terms found to be enriched in up-regulated (F) and down-regulated (G) genes from hOPC reporter cultures treated with astrocyte CM stimulated with or without TI. Genes were identified as differentially expressed if they had adjusted p-value less than 0.05 and Log2 Fold Changes values greater than 0.5 or less than –0.5, respectively. Statistical analyses reported in (B,C) are result of linear mixed effect models.