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Pseudo-spot data simulation overview: To simulate spot-level data from single cell level data, we created pseudo spots by partitioning the field of view into equal-sized squares. Counts are aggregated within each square, to create a pseudo-spot. Spot size is defined as the average number of cells in a pseudo-spot. We applied niche-DE to these lower-resolution datasets, and using the niche-DE results from the original high-resolution datasets as the gold standard, we computed the sensitivity and specificity of the niche genes found at each spot size.

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