Abc kit for color development

Colon tissues were fixed in 4% paraformaldehyde and embedded in paraffin. Tissue sections were prepared by the Translational Pathology Core Laboratory (University of California, Los Angeles). Sections were blocked and incubated with a rabbit polyclonal von Willebrand Factor (vWF) antibody (Millipore, Billerica, MA) overnight at 4°C. After washing, sections were incubated with donkey anti-rabbit IgG and slides, stained with an ABC kit for color development (Santa Cruz, CA) and photographed under the microscope and computerized. Image analysis of vWF-stained cells was performed using the Scion Image Software as we described^{8 (link)}. For histological scoring, sections were stained with H&E, photographed at multiple locations and analyzed by scoring specimens on a 0 to 10 scale for the following colitis parameters: mucosal integrity (0 to 6 scale), mucosal neutrophil infiltration (0 to 3 scale) and edema (0 to 1 scale)^{8 (link)}.

Colon tissues were fixed in 4% paraformaldehyde and embedded in paraffin. After incubation with blocking buffer, sections were incubated with a goat polyclonal anti-vimentin antibody (sc-7557, Santa Cruz biotechnology, Santa Cruz, CA, 1:50 dilution), a rabbit polyclonal anti-F4/80 antibody (sc-26643-R, Santa Cruz, 1:50 dilution) or a rabbit polyclonal anti-phospho-ERK antibody (#4370, Cell signaling, Danvers, MA, 1:50 dilution) overnight at 4°C. After washing, sections were incubated with donkey anti-goat IgG or bovine anti-rabbit IgG and slides were stained with an ABC kit for color development (Santa Cruz, sc-2018). Immunohistochemistry was assisted by Translational Pathology Core Laboratory (TPCL) of UCLA. Images were analyzed with a Zeiss AX10 microscope at magnification of 200×.