Fig. 4

CD8⁺ T cells promoted the differentiation of NSCs into astrocytes via the IFN-γ-Stat1 pathway in vitro. A qRT-PCR showing the expression of astrocyte markers (GFAP and Aldh1l1) in NSCs with or without a conditioned medium for 7 days. n = 5. **P < 0.01, ****P < 0.0001, one-way ANOVA with Tukey. B and C Immunofluorescence showing the percentage of astrocytes (GFAP⁺ cells) differentiated from NSCs with or without conditioned medium for 7 days. n = 35. ****P < 0.0001, one-way ANOVA with Tukey. Scale bar: 50 μm. D qRT-PCR showing the expression levels of GFAP and Aldh1l1 in NSCs with or without an anti-IFN-γ antibody for 7 days. n = 5. *P < 0.05, ****P < 0.0001, one-way ANOVA with Tukey. E and F Immunofluorescence staining showing the percentage of astrocytes differentiated from NSCs with or without an anti-IFN-γ antibody for 7 days. n = 35. ****P < 0.0001, one-way ANOVA with Tukey. Scale bar, 50 μm. G qRT-PCR showing the expression levels of GFAP and Aldh1l1 in NSCs with or without Stat1 siRNA for 7 days. n = 5. *P < 0.05, ****P < 0.0001, one-way ANOVA with Tukey. H and I Immunofluorescence staining showing the percentage of astrocytes differentiated from NSCs with or without Stat1 siRNA for 7 days. n = 35. ****P < 0.0001, one-way ANOVA with Tukey. Scale bar, 50 μm. J Visual representation of Stat1 ChIP-seq enrichment in the vicinity of the Aldh1l1 and GFAP locus using IGV software (V2.13.2). K Stat1 binding sites in the vicinity of the Aldh1l1 and GFAP locus. L Stat1 ChIP-qRT-PCR showing specific enrichment of the R2 and R3 regions upstream of the Aldh1l1 gene. R1 and R2: n = 5, R3: n = 6, *P < 0.05, one-way ANOVA with Tukey. M Stat1 ChIP-qRT-PCR showing specific enrichment of the R1 and R3 regions upstream of the GFAP gene. R1: n = 6, R2: n = 4, R3: n = 5, **P < 0.01, one-way ANOVA with Tukey