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The slides were washed with four changes of PBS, and incubated with a working dilution of biotin-labeled anti-mouse secondary antibody and avidin-biotin-peroxidase complex (ABC).
For cell culture assays, 10 mg/ml stock solutions were prepared in dimethyl sulphoxide (DMSO) and stored at -20 C. Working dilutions were prepared in assay medium immediately before use as described below.