Immunofluorescence Staining
The frozen sections of rat retina were washed, penetrated with Triton and blocked with goat serum blocking solution for 30 min. Sections were incubated with rabbit anti-rat SIRT1 polyclonal antibody (ab189494, 1 : 100; Abcam, Cambridge, UK) overnight, respectively. Next, sections were incubated with fluorescent second antibody for 1 h, stained by DAPI and sealed with glycerinum. The sections were finally observed under a confocal microscope to obtain the image of staining and images were analyzed by a confocal software.