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. Author manuscript; available in PMC: 2017 Jul 26.
Published in final edited form as: Nature. 2017 Jan 18;541(7638):481–487. doi: 10.1038/nature21029

Figure 4. Astrocyte-derived toxic factor promoting cell death.

Figure 4

Representative phase image showing death of purified retinal ganglion cells (RGCs) in culture (ethidium homodimer stain in red shows DNA in dead cells (a, 24 h quantification in c), and differentiated oligodendrocytes (b, 24 h quantification in d). e, Quantification of A1-induced cell death in human dopaminergic neurons (5 days). f, Western blot analysis for cleaved caspase-2 and -3 in RGCs treated with control and A1 ACM. g, retro-orbital optic nerve crushes (ONC) produced A1s in the retina. Intravitreal injection of neutralizing antibodies to Il-1α, TNFα, and C1q blocked A1 production. h, RBPMS (RNA-binding protein with multiple splicing, an RGC marker) immunostaining of whole-mount retinas showed decreased number of RGCs in ONC, rescued with neutralizing antibody treatment. Quantification is shown post ONC at 7 days (i), 14 days (j) using neutralizing antibodies, and at 7 days using Il1α−/−TNFα−/− and Il1α−/−TNFα−/− C1q−/− animals (k), and microglia-depleted (PLX-3397-treated) animals (l). * p < 0.05, one-way ANOVA. n = 8 in each instance. Error bars indicate s.e.m. Scale bar: 100 μm (a,b); 20 μm (k).