Fig. 1

VISTA is differentially expressed in distinct MS lesion stages and decreases in EAE. a Representative images of in situ expression of VISTA in distinct MS lesion stages. b Quantification of the area positive for VISTA in MS lesion stages compared to normal-appearing regions shown as log fold change. At least 3 images were taken from each lesion and the area positive for VISTA was measured. Statistical analysis performed was a Wilcoxon signed-rank test comparing lesions to normal-appearing areas with Benjamini–Hochberg correction for multiple comparisons (n = 5 for each lesion). (c) VISTA geometric mean fluorescence intensity (gMFI) on viable (DAPIneg) microglia (CD45intCD11BposLY6Cneg) during different stages of EAE including score 1(E1: early disease) score 4 (E4: peak disease), and chronic EAE (Ech), and unimmunized control (c) mice. Error bars indicate mean ± s.d. (n = 5–6). Statistical analysis performed was a one-way ANOVA with Dunnett’s correction for multiple comparisons. d Fold change in VISTA expression in primary neonatal mouse microglia 5 h after stimulation with LPS, Pam3CSK4, PolyI:C, myelin, IL1b, or TNFa, measured by RT-qPCR and normalized to Hprt1. Error bars indicate mean ± s.d. (n = 3–5). Statistical analysis performed was a one-way ANOVA with Dunnett’s correction for multiple comparisons. Boxes indicate median and lower and upper quartiles, whiskers show min/max, and outliers are indicated by points. ns = not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001