Anti gfp

Eyes were harvested and fixed for 1 h in 4% PFA. Following cryopreservation in 10%, 20%, and 30% sucrose at 4°C for 30 min, 2 h and overnight, respectively, eyes were embedded in Tissue-Tek OCT compound (Electron Microscopy Sciences) and sectioned at 10 μm using a cryostat. The following primary antibodies were incubated with tissue sections overnight at 4°C: anti-Iba1 (1:200, Abcam), anti-CD68 (1:250, Bio-Rad), anti-GFAP (1:500, Abcam), anti-DLK (1:100, Genetex), anti-Atf3 (1:200, Novus Biologicals), anti-GFP (1:500, Abcam), anti-Il22Ra1 (1:200, Bioss), anti-Rbpms (1:100; PhosphoSolutions), anti-βIII-tubulin antibody (1:500, Promega). Following primary antibody application and washing, tissue was subsequently incubated in AF594, AF546, AF647, or AF488 secondary antibodies (1:400; Invitrogen, Thermo Fisher) for 1 h. DAPI (1:1000; Invitrogen) was used to label nuclei. Images were captured using an Axio Imager M2 (Zeiss), a LSM 880 confocal (Zeiss), or a TCS SP8 confocal (Leica). To visualize AAV infection in wholemount retinas, tissue was stained with an anti-GFP (1:500, Abcam) antibody for 3 d at 4°C. Retinas were subsequently washed in PBS, incubated with an AF488 secondary antibody (1:400; Invitrogen) for 2 h at room temperature, and washed in PBS.

The following primary antibodies were used: anti-Krt14 (polyclonal chicken; 1:10,000; BioLegend), anti-GFP (green fluorescent protein; polyclonal rabbit; 1:1000; Abcam, RRID:AB_305564), anti-GFP (polyclonal goat; 1:1000; Abcam, RRID AB_305643), anti-GFP (polyclonal chicken; 1:500; Invitrogen, RRID:AB_2534023), anti-p63 (rabbit monoclonal; 1:1000; Abcam, RRID:AB_10971840), anti-Krt8 (rat monoclonal; 1:250; DSHB, RRID:AB_531826), anti-Entpd2 (rabbit; 1:4000; http://ectonucleotidases-ab.com, mN2-36Li6), anti-Gnat3 (goat polyclonal; 1:500; Novus Biologicals, NBP1-20926), anti-Snap25 (rabbit polyclonal; 1:500; Sigma-Aldrich, S9684), anti-Tas1r2 (rabbit; 1:500; Invitrogen, PA5-99935), anti-Lef1 (rabbit monoclonal; 1:100; Thermo Fisher Scientific, MA5-14966), anti-Sox2 (rabbit monoclonal; 1:200; Abcam ab92494).
The following secondary antibodies were used: anti-rabbit, anti-rat, anti-chicken, anti-goat conjugated to Alexa Fluor 488 (1:500; Jackson ImmunoResearch), to rhodamine Red-X (1:500; Jackson ImmunoResearch), or to Cy5 (1:1000; Jackson ImmunoResearch).