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. 2021 Jan 19;10:607349. doi: 10.3389/fonc.2020.607349

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Copyright © 2021 Kulasinghe, Taheri, O’Byrne, Hughes, Kenny and Punyadeera

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Tissue morphology (NanoString GeoMX DSP) for HNSCC tumors with 12 regions of interests (ROIs) per section, visualization markers and heatmap per tissue section. HNSCC tissue was profiled using four-color fluorescence of CD8 (red), CD3 (yellow), PanCK (green), and DNA (blue). Twelve 200 µm diameter circular regions of interest (ROIs) were selected and further profiled with a 40-plex oligo-antibody cocktail (individual ROIs below the tumor staining and counts for CD3, CD8, and PanCK cells per ROI). Unsupervised hierarchical clustering heatmaps of region-specific (tumor – green, Immune- red, Infiltrating edge with a mixed cell population – yellow) nCounter digital counts across all protein targets.