Immunohistochemistry staining of formalin-fixed and paraffin-embedded tissues was performed as previously described26 (link). After micro-sectioning at 4 μm thickness, the sections underwent heat-induced antigen retrieval for 3 min in EDTA buffer (pH 9.0; Dako, Carpinteria, CA). Sectioned slides were then incubated overnight with anti-human NIS (1:100 dilution; Santa Cruz Biotechnology, #sc-134515) at 4 °C. This was followed by incubation with HRP-labeled polymer-conjugated secondary antibodies against rabbit IgG (Dako) for 30 min at room temperature. A color reaction was induced using a ready-to-use 3,3′-diaminobenzidine substrate-chromogen solution (Dako) for 5 min, followed by washing with distilled water. Finally, sections were lightly counterstained with Mayer’s hematoxylin for 30 s before dehydration and mounting.
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