flow-sorted cells were collected from fresh tissue samples and processed for scRNA-seq in 156 sites from 41 patients .For each specimen with scRNA-seq data, site-matched FFPE tissue sections were used for whole-slide H&E staining and computational analysis .For each specimen with scRNA-seq data, site-matched FFPE tissue sections adjacent to the H&E section were stained by mpIF for major cell type and immunoregulatory markers .
and subjected to DNA extraction for bulk WGS. Genomic DNA was extracted using DNeasy Blood & Tissue kits and quantified on a Qubit 3 Fluorometer using the Qubit 1× dsDNA HS Assay kit .PBMCs were brought up to a volume of 15 ml in cold PBS, and DNA was isolated with the DNeasy Blood & Tissue kit according to the manufacturer’s protocol with 1 h of incubation at 55 °C for digestion. DNA was eluted in 0.5× buffer AE.
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