Anti mouse ep cam antibody

ICR mice (6/group) were treated with aerosol Aza at 2.5 or 75 mg/m² daily×7 days. One group of mice was treated with IT Aza at the maximum tolerated dose of 270 mg/m² as positive control for lung toxicity. The lungs were resected 7 days after the last aerosol administration. The lung toxicity was determined by pathological evaluation (3 lungs/group) using a previously described method [34] (link). Additionally, the viability of the airway epithelial cells was also determined (3 lungs/group). Briefly the lung tissue was digested with liberase (Roche) and filtered to obtain a single cell suspension, which was labeled with anti-mouse Ep-CAM antibody and fluorescence-conjugated goat anti-rat secondary antibody (Biolegend, San Diego, CA), and sorted by flow cytometry. The percentage of viable cells was determined by the exclusion of 4′, 6-diamidino-2-phenylindole staining. The systemic toxicity was determined by measuring blood cell counts as an indicator of myelosuppression (the major toxicity of IV Aza) [35] (link). Blood samples were taken at the time of euthanasia by cardiac puncture 7 days after last treatment. The total number of white blood cells (WBC) was determined after removal of red blood cells as previously described [32] (link).