Mbs2026249

Eyes from one animal were used for immunohistochemistry. Dissection and fixation were performed as previously described [13 (link)]. Hematoxylin and eosin staining was performed as described by Kiernan [52 (link)]. Anti-bodies used for staining included a polyclonal IgG antibody directed at IL-18 (MBS2026569, MyBioSource, San Diego, CA, USA), a polyclonal IgG antibody directed at S100A12 (MBS2026249, MyBioSource, San Diego, CA, USA), a polyclonal IgG antibody directed at ANXA1 (MBS2001804, MyBioSource, San Diego, CA, USA), a poly-clonal IgG antibody directed at fibronectin (ab23751, Abcam, Cambridge, UK), and a monoclonal IgG1 antibody directed at rhodopsin (ab190307, Abcam, Cambridge, UK). The antibodies were diluted in (1:200–1:800) in PBS + 0.3% Triton X100. The sections incubated overnight at 4 °C and were processed with EnVission (DakoCytomation) DAB. Controls were incubated with rabbit IgG_2b or irrelevant rabbit anti-bodies.

Western blotting was performed as previously described [13 (link)] using a primary monoclonal mouse anti-β-actin antibody 1:5000 (clone AC-15, Sigma-Aldrich, St. Louis, MO, USA), a primary polyclonal rabbit anti-IL-18 antibody 1:500 (MBS2026569, MyBioSource, San Diego, CA, USA), and a primary polyclonal rabbit anti-S100A12 antibody 1:100 (MBS2026249, MyBioSource, San Diego, CA, USA), diluted in 2.5% (w/v) skim milk blocking buffer. Log transformed densitometric data was used to perform a paired t-test.