IL-6 expression in colon tissues was inspected using a standard immunohistochemistry method. Immunostaining was done on serial sections as described previously by Zeng et al. [11 (link)]. Briefly, the tissues were fixed in 10% formaldehyde and embedded in paraffin and then cut into 4 μm sections. Tissue sections were, respectively, deparaffinized in xylene and rehydrated by grade alcohols. The tissue was incubated overnight at 4°C with primary antibody specific for IL-6 (Santa Cruz, CA, USA, dilution 1 : 200). The tissue sections were then incubated with biotin-labeled goat anti-mouse antibody (Sigma, USA) followed by exposure to avidin-peroxidase complex (Sigma, USA). Staining was developed with diaminobenzidine (DAB, Sigma) substrate and sections were counterstained with hematoxylin.
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