Cm dii red

Whole-mount retinas from P13 animals were permeabilised with 0.5% (wt/vol.) Triton-X100 at room temperature for 1 h. Overnight staining was with FITC/TRITC-labelled isolectin-B4 (1:50; Sigma-Aldrich) at 4°C. After PBS washes, retinas were flat-mounted in glycerol and micrographs were obtained with a fluorescence microscope (Lectin-FITC/ASC-Dil red staining; Leica BMR, Bensheim, Germany) or a confocal microscope (Lectin-TRITC/ASC-EGFP staining; Leica TCS SP2 confocal microscope, Leica, Wetzlar, Germany). ASCs presence was revealed through their pre-injection label, which was either CM-DiI-red (ThermoFisher, Waltham, MA, USA) or enhanced green fluorescent protein (EGFP) lentiviral tag.

The cell sheets from passage 3 c-kit+ CPC were formed using thermoresponsive polyNIPAM-coated 12-well UpCell™ plates (cat#Z688835, Sigma-Aldrich, USA). Cultured CPC were labeled by CM DiI red (cat#C7000, ThermoFisher Scientific, USA) and then incubated on thermoresponsive dishes at 37°C for 72 hours. The CM DiI-labeled CPC spontaneously detached from the dish surface following incubation at 20°C for 30 minutes, yielding a CPC sheet.