Freshly excised eye-balls were snap frozen in Tissue-Tek optimum cutting temperature compound (Sakura Finetechnical, Tokyo, Japan); frozen sections of 6-μm thickness were fixed in 4% paraformaldehyde for 15 min, permeabilised with 0.3% Triton X-100 for 5 min, and blocked with goat serum (zhongshan, Beijing, China) for 1 h. Tissue was incubated with primary antibody (anti-caspase-1; 1:500 dilution; Novus, UK) at 4 °C overnight. Tissue was then washed three times with phosphate-buffered saline (PBS) and then incubated with secondary antibody (green, Abcam, UK) for 1 h (1:500). For detection of cell nuclei, DAPI was added to the mounting medium. Tissues were imaged via fluorescence microscopy.

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