For flat-mount retina staining, fixed retinas were blocked overnight in 4°C with 1% bovine serum albumin and 0.3% Triton X-100 in phosphate-buffered saline (PBS). Retinas were conditioned with PBLEC buffer [1% Triton X-100, 1 mM CaCl2, 1 mM MgCl2, and 0.1 mM MnCl2 in PBS (pH 6.8)] before they were incubated overnight with a 1:25 Daylight 594–conjugated isolectin B4 (Vector Laboratories, Burlingame, CA, USA) in a PBLEC buffer at 4°C to stain for retinal vessels. After washing, the stained retinas were postfixed with 4% paraformaldehyde (PFA) for 5 min and washed again with PBS. Images of retinal vessels were taken with a confocal sSP5 microscope (Leica Microsystems, Germany).

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