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Fig. 5 A single intraocular dose of colchicine delays axon regeneration and makes axonal varicosities smaller and more abundant. (A) Representative image of GFP+ regenerating RGC axons 2 days postinjury after intraocular vehicle injection at 1 day postinjury (red circle, injury site). (B) Representative image of GFP+ regenerating RGC axons at 2 days postinjury after intraocular colchicine injection on 1 day postinjury (red circle, injury site). (C) Axon counts by the distance to the injured site with different surgery pins. Gray boxes represent P < 0.05 for vehicle versus colchicine post hoc comparison. (D) Representative image of GFP+ regenerating RGC axons and axonal varicosities (partially marked by arrowheads) 2 days postinjury after intraocular vehicle injection at 1 day postinjury. (E) Representative image of GFP+ regenerating RGC axons and axonal varicosities (partially marked by arrowheads) 2 days postinjury after intraocular colchicine injection at 1 day postinjury. (F) Varicosity density on axons increases after colchicine treatment. (G) Varicosity size decreases after colchicine treatment. Data in (C) are presented as mean ± SEM, ∼5 retinas from 3 animals were used in experiments, 2-way ANOVA with Bonferroni post hoc were used. For (F, G), ∼200 varicosities from ∼3 animals were measured in each group for violin plot, Student's t-test was used, ***P < 0.001. Scale bar: (A, B), 200 μm; (D, E), 50 μm. ANOVA, analysis of variance.