A Ventana BenchMark Ultra stainer (Ventana Medical Systems, Tucson, AZ, USA) was used to perform IHC on tissue sections with a thickness of 2 µm. A variety of antibodies were added, including epidermal growth factor receptor (EGFR; clone 3C6; Ventana Medical Systems), HER2 (clone 4B5; Ventana Medical Systems), HER3 (clone SP71; Abcam, Cambridge, UK), mTOR (clone 49F9; Cell Signaling Technology, Danvers, MA, USA), programmed death-ligand 1 (PD-L1; clone E1L3N; Cell Signaling Technology (until mid-2018; as of mid-2018, Nordic Biosite, Stockholm, Sweden, is using the BSR90 clone)), and PTEN (clone Y184; Abcam).
To assess the immunostaining intensity for the antigens EGFR, mTOR, PDGFRA, PDGFRB, and PTEN, a combinative semi-quantitative score was used. For a comprehensive description of the IHC, we refer to our previous work [9 (link)].
To assess the immunostaining intensity for the antigens EGFR, mTOR, PDGFRA, PDGFRB, and PTEN, a combinative semi-quantitative score was used. For a comprehensive description of the IHC, we refer to our previous work [9 (link)].
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