Figure 5 from An Integrated Spatial Multi-Omics Workflow for Sequential RNA and Protein Profiling in FFPE Tumor Tissue

<jats:p>&lt;p&gt;Multi-omic characterization of immune cell subsets within a lymphoid aggregate. &lt;b&gt;A,&lt;/b&gt; IMC image showing the structural organization of lymphoid cells in the immature TLS containing CD20 expressing B cells adjacent to tumor expressing keratin 8/18. Cells within this lymphoid aggregate are also seen to express Ki-67, a proliferation marker. &lt;b&gt;A′,&lt;/b&gt; Zoomed in inset shows the spatial positioning of B-cell aggregates and subcellular localization of Ki-67 within replicating cells. &lt;b&gt;A″,&lt;/b&gt; Coregistered transcriptomics and proteomics data reveal the expression of cytokine transcripts &lt;i&gt;CXCL9&lt;/i&gt;, &lt;i&gt;CXCL10&lt;/i&gt;, and &lt;i&gt;CXCL13&lt;/i&gt; in the vicinity of the immature TLS containing B cells. &lt;b&gt;B,&lt;/b&gt; IMC image demonstrating the spatial distribution of CD4 T cells, T cytotoxic cells (CD8), and memory T cells (CD45RO) in the immature TLS. &lt;b&gt;B′,&lt;/b&gt; Complex cellular organization of T and B cells can be detected across the entire lymphoid structure. &lt;b&gt;B″,&lt;/b&gt; Coregistered transcriptomics and proteomics data highlight the expression of &lt;i&gt;GZMK&lt;/i&gt; transcript on cytotoxic T cells. Expression of transcripts for checkpoint molecules &lt;i&gt;LAG3&lt;/i&gt; and &lt;i&gt;TIGIT&lt;/i&gt; is detected on cytotoxic T cells and other cells in this ROI. &lt;b&gt;C,&lt;/b&gt; IMC image showing the localized distribution of CD4 T cells in the developing TLS and its spatial relationship with the adjacent metastatic TME. &lt;b&gt;C′,&lt;/b&gt; IMC image showing the localization of Tregs expressing nuclear transcription factor FoxP3. &lt;b&gt;C″,&lt;/b&gt; Coregistered transcriptomics and proteomics data show expression of corresponding transcripts for &lt;i&gt;FOXP3&lt;/i&gt; as well as &lt;i&gt;CTLA4&lt;/i&gt; and &lt;i&gt;TNFRSF9&lt;/i&gt; on Tregs. Scale bars, 200 μm.&lt;/p&gt;</jats:p>