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. 2023 May 11;14(5):319. doi: 10.1038/s41419-023-05834-9

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2023

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Fig. 4 — A H&E-stained section (left) fused with a serial fluorescent image (right). H&E sections were evaluated by a pathologist who defined areas of necrosis (purple), viable tumor (green), and stroma (orange). The %NOS2/COX2-expressing cells in the entire tumor as well as tumor and stroma, are shown. B Areas in the spatial distribution highlighted by blue boxes labeled 1 (top), 2 (middle), and 3 (bottom) reflect a progression from (1) inflamed regions of stroma-restricted CD8⁺ T cells to (2) cold regions with reduced stroma-restricted CD8⁺ T cells and (3) cold immune desert tumor core regions lacking CD8⁺ T cells. Spatial analyses of NOS2/COX2 expression in these boxed areas as well as CD8⁺ T cells or IFNγ, are shown at 50 μm magnification. Registered images in the inflamed region designated in box 1 shows C stroma-restricted CD8⁺ T cells (cyan) or D IFNγ expression (cyan) near NOS2-expressing cells (red). Analyses of cold regions designated in box 2 show high COX2 expression and low NOS2 expression with E limited CD8⁺ T cells and F limited IFNγ. Analyses of immune desert tumor core regions associated with box 3 show high tumor COX2 expression with abated levels of G CD8 + T cells and H abated IFNγ expression.