Fig. 6: CSPG-exposed Th17 cells are more toxic to oligodendrocytes. | Nature Communications

Fig. 6: CSPG-exposed Th17 cells are more toxic to oligodendrocytes.

From: Versican promotes T helper 17 cytotoxic inflammation and impedes oligodendrocyte precursor cell remyelination

Fig. 6

a 2D2 T cells were isolated and activated with MOG-loaded dendritic cells and differentiated to Th17 subset in ECM-coated plates and they were then added to OPCs for 24 h. Schematic diagram of experiment procedure is shown (Created with BioRender.com). b, c Live imaging of labeled OPC-T cell co-culture was performed with PI in the culture medium, and images at different time points are displayed. b Representative images of OPCs (green); PBS-, CSPG- or versican-V1-exposed Th17 cells (red); and cellular PI (white) are provided. To rule out a direct effect of CSPG in mediating death of OPCs, CSPGs (10 μg/ml) was also added to the culture medium of OPCs (CSPG control). c Bar graph showing the proportion of PI labeled OPCs after 12 h, n = 9 replicates over three separate experiments. Data are presented as mean ± SEM. One-way ANOVA - Bonferroni post hoc; significance indicated as *p = 0.03, **p = 0.005, ***p < 0.001. d, e In a separate co-culture experiment, cells were not labeled with cell trackers and instead were stained for O4 (green) and MBP (red) markers after 24 h. OPC cell number or process outgrowth were measured using ImageXpress. d Representative images from OPC co-culture with PBS (Control)-, CSPG- or versican-V1-exposed Th17 cells and e bar graph comparing the fold change of O4+ OPCs and mature MBP+ oligodendrocytes between PBS-, CSPG- or versican-V1-exposed Th17 cells after 24 or 72 h; n = 10 replicates over three separate experiments. Data are presented as mean ± SEM. Two-way ANOVA - Bonferroni post hoc; significance indicated as ***p < 0.001. f Quantification of OPC process outgrowth following co-culture with Th17 cells exposed to different ECM proteins, n = 10 replicates over three separate experiments. Data are presented as mean ± SEM. One-way ANOVA- Bonferroni post hoc; significance indicated as ****p < 0.001. For each replicate, data from twelve images (FOVs) were averaged to a single data point. Source data are provided in the source data file.

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