Temporal phosphorylation changes were assessed by Western blot. For our experiments evaluating sarpogrelate without LE, retinas were collected 5 minutes, 20 minutes, 30 minutes, 1 hour 20 minutes, 6 hours, 12 hours, 24 hours, and 48 hours post sarpogrelate or saline injection. For our experiments evaluating sarpogrelate-mediated neuroprotection with LE, retinas were collected immediately after 20 minutes of LE, 1 hour of LE; or at 20 minutes, 1 hour, 3 hours, 7 hours, 24 hours, 48 hours, or 96 hours post LE. Retinas were harvested and sonicated in phosphate-buffered saline (1X PBS) containing protease and phosphatase inhibitors (Thermo Scientific). Loaded samples (200 μg) were separated on a 12% acrylamide gel and transferred onto a membrane overnight at 4°. The next day, membranes were incubated in Odyssey infrared imaging system blocking buffer (Li-Cor, Lincoln, NE, USA) for 1 hour. Blots were then labeled with the following rabbit polyclonal antibodies diluted at 1:1000: phospho-PKCβ Ser661 (A8169; Assay Biotechnology [AB], Sunnyvale, CA, USA), phospho-PKCα/βII Thr638/641 (9375; Cell Signaling Technology [CST], Danvers, MA, USA), phospho-MEK1/2 Ser217/221 (9121; CST), MEK1/2 (9122; CST), phospho-p44/42 MAPK (pERK1/2) Thr202/Tyr204 (9101; CST), p44/42 MAPK (ERK1/2) (9102; CST), phospho-PDK1 Ser241 (3438; CST), phospho-Akt Ser473 (9271; CST), Akt (9272; CST), phospho-CREB Ser133 (A7053; AB), B-cell lymphoma 2 (Bcl2, 2870; CST), and phospho-Fos Thr232 (A8226; AB). After overnight incubation in primary antibodies, blots were incubated in a goat anti-rabbit IRDye 680RD secondary antibody (Li-Cor) diluted at 1:15000 for 1 hour at room temperature. Finally, blots were imaged with an infrared imaging system (Odyssey, Odyssey 2.1 software; Li-Cor). As an internal control for protein loading, membranes were also probed with GAPDH (2118; CST) or eIF4E (9742; CST). Using optical density measurements obtained with Odyssey software, each sample was normalized to the loading control. Then experimental groups were compared to control groups to determine fold changes in phosphorylation due to LE or sarpogrelate treatment. The experiments were performed independently two times, two to three animals per group.