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. 2022 Jul 22;146(2):749–766. doi: 10.1093/brain/awac266

Figure 6.

Figure 6

RNAscope for male pain-associated genes TNF, OSM, IL1B and IL32. (A) RNAscope TNF expression (red) overlaid with TRPV1 expression (green), AIF1 (white) and DAPI (blue) in pseudocolour. (B) Schema showing how TNF-alpha signalling cascades overlap with other signalling pathways, based on existing literature (icons from BioRender). (C) TNF, OSM, IL1B and IL32 quantile plots show the shift in abundance between male pain and non-pain subcohorts, while the bar plots show the RNA-seq abundance for the patient DRGs that were queried by RNAscope assay. (D) RNAscope OSM, IL1B and IL32 expression (red) overlaid with TRPV1 expression (green), AIF1 (white) and DAPI (blue) in pseudocolour. Scale bar = 20 μm, large globular structures are considered to be lipofuscin. Yellow arrows point out cells with overlap of expression between the gene of interest (red) and AIF1 (white), cyan arrows point out overlap of expression between the gene of interest (red) and TRPV1 (green), respectively, in the zoomed-in images, and pink arrows point to lipofuscin. (A and D) In all micrographs, wide field views and zoomed-in views on single neurons and surrounding cells are shown to display overall signal distribution, and colocalization of signal with specific neuronal and macrophage cell markers for each RNAscope probe.