Staining was performed using an automated IHC system on formalin-fixed and paraffin-embedded tissue of representative sections that were cut into 2 μm thick samples according to the protocol used in a study of the Institute of Pathology and Molecular Pathology, University Hospital Zurich, using a Ventana Benchmark XT automated staining system (Roche Tissue Diagnostics, Basel, Switzerland) [29 (link)]. The samples were loaded either in the Bond-Max system (Leica Microsystems, Wetzlar, Germany) or BenchMark Ultra system (Roche Tissue Diagnostics, Basel, Switzerland).
The following antigenes were targeted with antibodies: MIF-1 (Abcam, Cambridge, UK), CXCR2 (Thermo Fisher, Waltham, MA, USA) and CXCR4 (Abcam, Cambridge, UK). MIF-2 and CD74 antibodies were provided by the Bernhagen lab, 81377 Munich, Germany.
The staining assessment was performed according to the study of He et al. [30 (link)]; it included the Remmele score, which is calculated by multiplying the intensity and percentage of staining. Expression was considered negative or low when the score ranged from 0 to 2 and positive or strong when the score was >2.
The following antigenes were targeted with antibodies: MIF-1 (Abcam, Cambridge, UK), CXCR2 (Thermo Fisher, Waltham, MA, USA) and CXCR4 (Abcam, Cambridge, UK). MIF-2 and CD74 antibodies were provided by the Bernhagen lab, 81377 Munich, Germany.
The staining assessment was performed according to the study of He et al. [30 (link)]; it included the Remmele score, which is calculated by multiplying the intensity and percentage of staining. Expression was considered negative or low when the score ranged from 0 to 2 and positive or strong when the score was >2.
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