Goat anti mouse endocan antibody

Mice with OIR and CNV or age-matched controls were euthanized. Their eyes were enucleated and rapidly frozen in embedding medium (Sakura Finetek, Torrance, CA, USA) prior to sectioning. Sections (10 lm) were thawed, air-dried and fixed in 4% paraformaldehyde at room temperature for 10 minutes. These sections were incubated with blocking buffer (PBS containing 10% FBS) for 1 hour, and were then subsequently with goat anti-mouse endocan antibody (R&D Systems) for 2 hours. They were then incubated with AlexaFluor 555 donkey anti-goat secondary antibody (1:500; Invitrogen, Carlsbad, CA, USA) and isolectin for 1 hour at room temperature in the dark. The nuclei were stained with DAPI (Beyotime Biotechnology, Shanghai, China) for 5 minutes. The sections were washed with PBS between these incubations. Finally, the sections were mounted with fluorescence mounting medium, and examined and captured with a fluorescence microscope.

Fresh isolated eye tissues and cells were lysed, quantified, blotted and developed as previously described. 18 The following primary antibodies were used for the purpose of this experiment: goat anti-mouse endocan antibody (0.1 lg/mL; R&D Systems); rabbit anti-matrix metalloproteinase (MMP)-2 antibody (1:1000; Cell Signaling Technology); MMP-9 antibody (1:1000; Cell Signaling Technology); rabbit anti-PIGF antibody (1:1000; Abcam, Cambridge, UK); goat anti-VEGF antibody (1:1000; Abcam); rabbit anti-VEGFR1 antibody (1:1000; Abcam); rabbit anti-VEGFR2 antibody (Cell Signaling Technology); rabbit anti-P38 antibody (1:1000; Cell Signaling Technology); rabbit anti-phospho (p)-P38 antibody (1:1000; Cell Signaling Technology); rabbit anti-extracellular signal-regulated kinase (ERK) antibody (1:1000; Cell Signaling Technology); rabbit anti-p-ERK (1:1000; Cell Signaling Technology); rabbit anti-c-Jun Nterminal kinases (JNK) antibody (1:1000; Cell Signaling Technology) and rabbit anti-p-JNK antibody (1:1000; Cell Signaling