Anti mcpip 1

Mouse corneal samples were lysed with RIPA buffer. The lysates were centrifuged to obtain supernatant. Protein concentration was determined by BCA assay. For Western blot analysis, the protein samples were separated by SDS-PAGE and electrically transferred onto nitrocellulose membranes (Bio-Rad Laboratories, Hercules, CA). The membranes were blocked with 5% milk and subsequently incubated with primary and secondary antibodies. Signals were visualized using SuperSignal West Pico chemiluminescent substrate (Thermo Scientific, Pittsburgh, PA). β-actin was used as the loading control. Quantification of protein levels was based on the densitometry of blots by using the software Carestream MI SE (Informer Technologies, Rochester, NY). The antibodies used included: anti-IL-23, anti-IL-23R, anti-IL-17A, anti-IL-17RC, anti-MCPIP-1, anti-Osteoprotegerin (R&D), and anti-β-actin (A1978; Sigma-Aldrich). Enzyme-linked immunosorbent assay (IL-23; R&D) and protein array (Proteome Profiler Array Mouse XL Cytokine Array Kit; R&D) were performed following manufacturer’s protocols.