Fig. 5

a Top 3 identified pathways enriched in the optic nerves of Nf1^flox/flox; hGFAP-Cre mice after optic nerve crush (ON-CR) relative to sham controls following filtering of differentially expressed transcripts from bulk RNA sequencing. b Increased IL-1β RNA expression (qPCR) is observed in the optic nerves of 12-week-old Nf1^flox/flox; hGFAP-Cre mice following ON-CR at 6 weeks of age relative to sham controls (n = 4). c Representative IL-1β immunostaining in the optic nerves of Nf1^flox/flox; hGFAP-Cre mice following ON-CR relative to sham control mice. d RNAscope (in situ RNA hybridization) demonstrates that oligodendrocytes (Olig2⁺ cells) express IL-1β. e Immunostaining reveals that the majority of the Olig2⁺ cells co-label with CC1, but not with NG2. f Increasing IL-1β concentrations (0–75 ng/ml) increases microglia Ccl5 protein expression in vitro. g IL-1β-induced microglial Ccl5 production is attenuated by 10 mg/kg NFκB inhibitor (NFκB-IN; Caffeic acid phenethyl ester) treatment in vitro (n = 4). h Anti-IL1β neutralizing antibody treatment (1 mg/ml) of Nf1^flox/flox; hGFAP-Cre mice immediately after ON-CR at 6 weeks of age results in decreased i optic nerve proliferation (%Ki67⁺ cells; n = 5) and j Ccl5 mRNA expression (qPCR; n = 3) when analyzed at 12 weeks of age compared to IgG-treated (1 mg/ml) controls. Data are presented as the means ± SEM. Scale bars: c, d, e, i 50 µm. b, c, e, i, j. Two-tailed Student’s t test; f, g One-way ANOVA with Bonferroni post-test correction