Fig. 6

ET-1 colocalizes with reactive astrocytes and myeloid cells during progressive EAE in GFAPγR1Δ mice. a Representative confocal images (Z stack) of spinal cords from WT and GFAPγR1Δ mice, treated with isotype control or anti-TNF mAb, during chronic (d30) EAE stained for ET-1 (red), DAPI (blue), and either GFAP to identify astrocytes (left, green), or CD11b to identify macrophages/microglia (right, green), Scale bars, 50 μm. Images are representative of three to four separate fields from two to three mice per group. The distribution of ET-1 reactivity among astrocytes (b) or myeloid cells (c) was quantified by dividing the dually ET-1⁺ GFAP⁺ staining area by the total ET-1⁺ area per field or the ET-1⁺ CD11b⁺ staining area by the total ET-1⁺ area per field, respectively, in the fields described under panel a. Values are plotted as percentages of ET-1⁺ GFAP⁺ or ET-1⁺ CD11b⁺ reactivity within total ET-1 reactivity in panels b and c, respectively. Conversely, the percent of astrocytes (d) or myeloid cells (e) expressing ET-1 was assessed by dividing the ET-1⁺ GFAP⁺ or ET-1⁺ CD11b⁺ dual staining area by the e total GFAP⁺ or CD11b⁺ area within demyelinated lesions. Data represent mean ± SEM. P values were determined by one-way ANOVA on rank-sum test. GFAPγR1Δ in all panels represents GFAPγR1Δ mice treated with isotype control mAb