*2.1. RNA-Sequencing Comparison and Identification of Compounds via the Comparative Toxicogenomics Database*

Previous results of RNA-sequencing from three different animal models (mouse controlled optic nerve crush [3], DBA/2J mouse model of glaucoma [8], and rat optic nerve transection [4]) were examined. Matrices of DE genes were analyzed at a false discovery rate (FDR, *q*) of 0.05 (0.10 for the mouse controlled optic nerve crush, as reported in the original study). Gene lists were compared in a three-way analysis in R showing a number of shared DE genes between each model. Pathway analysis was performed (Ingenuity pathway analysis, Qiagen, Hilden, Germany) on shared DE genes between comparisons. The shared DE gene list common to all three models was queried in the Comparative Toxicogenomics Database (CTD), which compiles known gene/protein interactions with chemicals based on published data, and compounds targeting a significant proportion of the identified DE genes (>50% occurrence) were identified. Compounds for neuroprotective testing were selected based on previous neuroprotective literature support and novelty to glaucoma.

#### *2.2. Animal Strain and Husbandry*

All breeding and experimental procedures were performed in accordance with the Association for Research for Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Research. Individual study protocols were approved by Stockholm's Committee for Ethical Animal Research (10389-2018). All mice were housed and fed in a 12 h light/12 h dark cycle with food and drinking water available *ad libitum*. C57BL/6J and B6.Cg-Tg(Thy1-CFP)23Jrs/J (JAX stock number #003710; CFP + RGCs) mouse strains were purchased from The Jackson Laboratory (Bar Harbor, ME, USA) and bred and maintained in-house. All mice were used at 12–20 weeks of age.
