Paraffin-embedded tissue sections (4 μm) were deparaffinized, subjected to antigen retrieval in hot 10 mM sodium citrate buffer, and then blocked with BSA solution for 1 h at RT. They were incubated with primary antibodies overnight at 4 °C, washed with PBS, incubated with secondary antibodies for 30 min at RT, and developed with DAB reagent (Dako). Sections were counterstained with hematoxylin, dehydrated, and then coverslips were mounted with DPX (Sigma) and slides observed and photographed under the light microscope CX40 Olympus, equipped with the Altra-20 digital camera51 (link).
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