Rabbit anti rbpms gtx118619

Hypoxia was assessed using pimonidazole HCl (Hypoxyprobe) according to manufacturer’s instructions. Ninety minutes prior to euthanasia, mice were given an intraperitoneal injection of (60 mg/kg) pimonidazole HCl diluted in sterile 1X PBS. Mice were then transcardially perfused with 20 mL saline solution and 20 mL 4% paraformaldehyde. Eyes were harvested and incubated in 4% paraformaldehyde for 30 min. Eyes were stored in 1X PBS until dissection. Hypoxia was detected using an antibody against pimonidazole (Table 1).Table 1

Summary of antibodies.

	Concentration	Company
Primary antibody (catalog number)
Rabbit anti-cCASP3 (AF835)	1:1000	R&D Systems
Goat anti-CD31 (AF3628)	1:1000	R&D Systems
Goat anti-ChAT (AB144P)	1:200	Millipore
Mouse anti-JUN (BDB610327)	1:250	Fisher Scientific
Rabbit anti-pimonidazole (PAb2627AP)	1:200	Hypoxyprobe
Rabbit anti-pJUN (9261S)	1:250	Cell Signaling
Rabbit anti-RBPMS (GTX118619)	1:250	GeneTex
Guinea pig anti-RBPMS (1832-RBPMS)	1:250	PhosphoSolutions
Goat anti-SOX2 (sc-17320)	1:200	Santa Cruz
Secondary antibody (catalog number)
Donkey anti-rabbit (A31572, A-21206)	1:1000	ThermoFisher
Donkey anti-guinea pig (706–605–148)	1:1000	Jackson ImmunoResearch
Donkey anti-goat (A21447)	1:1000	ThermoFisher
Donkey anti-mouse (A31570)	1:1000	ThermoFisher

As previously described^{38 (link),39} , eyes were harvested and fixed in 4% PFA in 1× PBS for 90 min. Retinas were dissected free from the optic cup and blocked in 10% horse serum, 0.4% Triton™ X-100 (Fisher scientific, 9002-93-1) in 1× PBS overnight at 4 °C. Retinas were then incubated at 4 °C for 3 days in primary antibodies (Table 1) diluted in 10% horse serum, 0.4% Triton™ X-100 in 1× PBS. Retinas were then washed and incubated for 24 h at 4 °C in secondary antibodies (Table 1) diluted in 1× PBS. Retinas were washed and mounted on microscope slides ganglion cell layer-up in Flourogel in TRIS buffer (Electron Microscopy Sciences, 17985-11).Table 1

Summary of antibodies

	Concentration	Company
Primary antibody (catalog #)
Rabbit anti-cCASP3 (AF835)	1:1000	R&D Systems
Rabbit anti-RBPMS (GTX118619)	1:250	GeneTex
Guinea pig anti-RBPMS (1832-RBPMS)	1:250	PhosphoSolutions
Secondary antibody (catalog #)
Donkey anti-rabbit 555 (A31572)	1:1000	ThermoFisher
Donkey anti-guinea pig 647 (706-605-148)	1:1000	Jackson ImmunoResearch

cCASP3 cleaved caspase 3, RBPMS RNA binding protein, mRNA processing factor