Figure 6. Minocycline treatment does not neuroprotect RGCs following rNAION induction and cytokine analysis. A: Cytokine analyses of vehicle- and minocycline-treated anterior optic nerves (ONs) 3 days post-rNAION induction. Five anterior ON segments were pooled from each condition and compared to the uninduced contralateral eye (vehicle-treated rodent nonarteritic anterior ischemic optic neuropathy (rNAION) induced against contralateral naïve: White bars. Minocycline-treated rNAION induced against contralateral eye: Black bars). Assays shown are all individual analyses with expression statistically significantly above baseline. Low-level (expression (1–10 pg/ml; graph shown on the left) and high-level (100–900 pg/ml; graph shown on the right) values were obtained. Cytokine expression rose after rNAION induction compared with uninduced nerves, for M1-associated cytokines (TNF-α, IL-1β, and IFNγ) and M2-associated cyto- and chemokines (G-CSF, IL-4, and IL-13). B–D: Representative images of Brn3a stained retinal ganglion cells are shown. Whole mount retinas from B: naïve, C: 30 days post-rNAION vehicle, and D: 30 days post-rNAION minocycline-treated retinas. Panels show global retinal ganglion cell (RGC) distribution using 4X magnification and direct RGC identification using 60X magnification inserts. E: Stereological analysis of RGC counts per square of retina for naïve (white bar) and vehicle- (black bar) and minocycline-treated (hatched bar) retinas 30 day post-rNAION induction. There is a slight difference in residual RGC numbers when vehicle- and minocycline-treated rNAION-induced eyes are compared, but this is not statistically significant (ANOVA; f-ratio = 2.63798, p=0.09).