Fig. 7

Pro-inflammatory cytokines associated with activated microglia and neurotoxic astrocytes are upregulated in EAE optic nerve. a Gene expression of pro-inflammatory cytokines, TNF-α, IL-1α as well as C1q in the optic nerve of EAE mice (n16) at peak stage using qPCR. b Astrocyte A1-specific and A2-specific transcripts expression in the optic nerve of EAE mice as previously published. c Astrocyte A1-specific and A2-specific transcripts expression in the purified astrocyte from hindbrain of EAE mice (n = 16). d GFAP, IBA1 and C3 costaining in the optic nerve of EAE mouse. Immunofluorescence staining further confirmed that increased C3 was predominantly expressed in astrocytes and not microglia. e Quantification of cells double positive for C3 and GFAP staining (n = 8 for CFA; n = 11 for EAE). f GFAP and PSMB8 co-staining in the optic nerve of EAE mouse. Yellow arrowhead indicated GFAP+/PSMB8+ cell; red arrowhead indicated GFAP+/PSMB8- cell; white arrowhead indicated MAC2+/PSMB8- cell. g Quantification of cells double positive for GFAP and PSMB8 staining (n = 8 for CFA; n = 11 for EAE). Boxed regions from each image were magnified as shown in figures. White dashed lines delineate the ROI. Quantification was represented as mean ± SEM. Significance was determined by two-tailed, unpaired Student’s t-test with P < 0.05 consider as significant. * P ≤ 0.05, *** P ≤ 0.001. Scale bar =50 μm