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. 2021 Aug 19;11:16839. doi: 10.1038/s41598-021-96107-y

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Axonal counts in PPD-stained ON semi-thin sections and RBPMS⁺ RGC in retinal cryosections in the “pre-blast” injection study. (A) There were increased total degenerating axons in the ON after b-ITON compared to sham (P < 0.0001, ANOVA) but not in b-ITON eyes intravitreally injected with siCASP2 or siEGFP control (P < 0.001 for both siCASP2 and siCASP2 compared to b-ITON, post-hoc Tukey). The number of degenerating axons was lower in siCASP2 treated eyes compared to siEGFP controls, indicating a potential axon-protective effect of caspase-2 knockdown (P = 0.0170, post hoc Tukey). (B) There were no differences in intact axons between any groups (P = 0.214, ANOVA). (C) There was weak evidence of differences in ON axon density between sham and b-ITON groups (P = 0.1099, ANOVA). (D) Representative PPD and toluidine blue-stained resin semi-thin ON sections, with degenerating axons (arrowheads). (E) There were no differences in ON area between groups (P = 0.7244, ANOVA). (F) Representative IHC images showing RBPMS⁺ RGC in the GCL on retinal cryosections. (G) There was weak evidence of a difference in the number of RBPMS⁺ RGC after b-ITON with “pre-blast” intravitreal injections (P = 0.1364, ANOVA). Scale bar in (D) = 20 μm and (E) = 100 μm. Error bars represent mean ± SEM.