Figure 5.

Optic glioma formation and growth is partly determined by the effect of the germline Nf1 gene mutation on microglia number and function. (A) Iba1 immunostaining demonstrates increased numbers of microglia in the optic nerves of neo^CKO (3.6-fold) and R681X^CKO (7.1-fold) mice relative to controls (CTL). Whereas R681X^CKO mice have more microglia than neo^CKO mice (2-fold), no increase in microglia numbers was detected in G848R^CKO mice relative to controls (n = 5 mice per genotype). (B) Increased percentages of pJNK⁺ cells were observed in neo^CKO (4.66-fold) and R681X^CKO (11.28-fold) optic nerves relative to control optic nerves (n = 4 mice per genotype). (C) Immunofluorescence revealing pJNK staining in Iba1⁺ cells (microglia) within the optic nerves of R681X^CKO mice. White arrowheads depict two independent cells positively labeled with both pJNK and Iba1. (D) Increased percentages of CCL5⁺ cells were observed in neo^CKO (14.56-fold) and R681X^CKO (27.37-fold) optic nerves compared with control mice (n = 5 mice per genotype). (E) A greater percentage of pAkt^Thr308-immunoreactive cells was found in R681X^CKO mice relative to control (4.68-fold), and neo^CKO (2.88-fold) optic nerves (n = 4 mice per genotype). Scale bars: 100 µm. All data are represented as means ± s.e.m. (***P < 0.001; **P < 0.01; *P < 0.05; one-way ANOVA with Bonferroni post-test).