Fig. 5

Expression of various astrocyte genes and signaling pathways. A Functional ATP assay measuring ATP levels in optic nerve head tissues from control and 30 days microbead injected animals. MB = microbead injected. N = 4 for each group. Mean ± SEM. * p < 0.05. B Mitochondrial biogenesis in optic nerve head tissues from control and 30 days microbead injected animals. MB = microbead injected. N = 4 for each group. Mean ± SEM. ** p < 0.01. C Changes in the proportion of cellular ATP derived from glycolysis versus oxidative phosphorylation in response to elevated IOP within optic nerve head tissues. Oligomycin was used to inhibit ATP synthesis by oxidative phosphorylation. N = 4 for each group. Mean ± SEM. *** p < 0.01. D Reactive oxygen species was measured using DHE (dihydroethidium) labeling of live unfixed longitudinal sections of the optic nerve. E Log2 fold change values for various astrocyte genes, as determined in the differential expression analysis of the 3 independent time point pairwise comparisons. Astericks indicate significant differences between the 2 groups (adjusted p-value < 0.05). F Log2 fold change values for reactive astrocytes genes, as determined in the differential expression analysis of the 3 independent time point pairwise comparisons. Astericks indicate significant differences between the 2 groups (adjusted p-value < 0.05). G Gene set enrichment analysis on the 3 time point pairwise comparisons against a user defined subset of the GO, KEGG and Reactome pathway, focusing on key reactivity and injury signaling pathways: nuclear factor kappa B (NFkB), calcineurin-nuclear factor activating of T-cells (CaN-NFAT), mitogen-activated protein kinase (MAPK), janus kinases/signal transducer and activator of transcription (JAK/STAT), Wnt/b-catenin, and sonic hedgehog (adjusted p-value < 0.05). NES = normalized enrichment score