Immunofluorescence Analysis of Mouse Retina

Immunofluorescence analysis was performed with mouse retinal sections. Cryosections were prepared as described previously.²¹ (link) After fixation and blocking, primary antibodies used for staining were rat anti-HA (1:500, AF0039, Beyotime), chicken anti-GFP (1:500, ab13970, Abcam, Cambridge, MA), rabbit anti-Rhodopsin (RHO) (1:500, 1D4, Santa Cruz Biotechnology, Santa Cruz, CA), and rabbit anti-cone arrestin (1:500, AB15282, Merck Millipore, Billerica, MA). The samples were washed and then stained at room temperature for 1 hour with Alexa Fluor 594- and 488-conjugated secondary antibodies (1:1000, Proteintech). As for the quantification of RHO and cone arrestin, six sections each with six random fields were captured and measured by a masked observer.