The TMAs and whole slides were cut into 4 µm thick slides and MAB against DLL3 (Ventana, clone SP347, ready to use) was used, which was proven to be specific for DLL3-expression elsewhere (16, 17). IHC was performed with an OptiView Ventana System using the OptiView DAB IHC Detection Kit and the Cell Conditioning 1 pretreatment solution (CC1) according to the manufacturer’s protocol (All Ventana). Pretreatment with the CC1 solution was for 80 min at 100°C, primary body incubation for 32 min at 36°C, linker component for 8 min at room temperature followed by HRP substrate for 8 min at room temperature. Slides were counterstained with hematoxylin. As specified by the manufacturer, staining reaction is specific when seen membranous or cytoplasmic. As negative control kidney and liver tissue were used. As positive control slide with non-small lung cancer was parallel stained showing a positive signal for DLL3.

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