Corneas and cells were ground and lysed in radioimmunoprecipitation assay (Beijing Solarbio Science and Technology Co., Ltd., Beijing, China) lysis buffer containing 1-mM phenylmethanesulfonyl fluoride (Beijing Solarbio) for 2 hours. They were then centrifuged at 4°C, 12,000 rpm, for 10 minutes. The supernatant was collected and tested for protein concentration. After SDS sample buffer was added, followed by boiling, total protein was separated on 12% or 15% acrylamide SDS-PAGE and transferred onto polyvinylidene difluoride membrane (Beijing Solarbio). The membranes were blocked with Western Blocking Buffer (Beyotime Biotechnology, Shanghai, China) at room temperature for 2 hours and were then incubated with primary antibody to IL-36α (1:1000; R&;D Systems), IL-1β (1:1000; Abcam, Cambridge, UK), IL-6 (1:1000; Abcam), TNF-α (1:1000; Abcam), phosphorylated (p) NF-κB (1:1000; CST, Wuhan, China), total NF-κB (1:1000; CST), glyceraldehyde 3-phosphate dehydrogenase (1:2000; Elabscience, Wuhan, China), β-actin (1:2000; Elabscience), or β-tublin (1:2000; Elabscience) at 4°C overnight. After washing in PBS containing 0.05% TWEEN 20 (Bio-Rad, Hercules, CA, USA) for three times, the membranes were incubated with corresponding peroxidase-conjugated secondary antibodies (1:5000; Elabscience) at room temperature for 1 hour. The blots were then developed by using chemiluminescence (Thermo Fisher Scientific, Waltham, MA, USA).