Controlled optic nerve crush (CONC) was performed as previously described1 (link)3 (link). Mice were anaesthetized with a mix of ketamine and xylazine. The optic nerve was crushed just behind the eye for approximately 4 seconds using self-closing forceps (Roboz RS-5027). Unmanipulated contralateral eyes or contralateral eyes that had a sham surgery performed (no crush of the optic nerve) were used as control eyes. All CONC experiments were performed on B6.Bim mice. DBA/2J mice were used as a glaucoma model. The null allele of Bim was backcrossed into DBA/2J mice for 10 generations and then intercrossed. The TonoLab (Colonial Medical Supply, Franconia, NH) was used to record IOP in D2.Bim mice. Mice were anaesthetized with a ketamine xylazine mix and IOP was recorded per manufacturers instructions between two and five minutes after administration of anesthetic. For determining the level of glaucomatous optic nerve damage, nerves were processed and stained with paraphenylenediamine (PPD) as previously described3 (link)8 (link)29 (link) except that nerves were embedded in Technovit 7100 and 2 μm sections were cut and stained. Nerves were graded using a validated grading scale as previously described3 (link)8 (link)29 (link). The grading scale places eyes into three categories: no or early, less than 5% of the axons are thought to be damaged or lost, a number that is consistent with age-related damage; moderate, many damaged axons throughout the nerve averaging about 30% of the axons judged to be damaged or lost, often there is localized signs of gliosis; severe, greater than 50% of the axons are judged to be damaged or lost and often signs of large areas of glial scarring. For plastic sections of retinas, eyes were processed and cut as previously described1 (link).