For each sample, two FFPE scrolls of 25 µm were prepared and processed according to the "Isolation of Cells from FFPE Tissue Sections for Chromium Fixed RNA Profiling" protocol (CG000632, 10x Genomics), employing pestle-based dissociation. After dissociation and deparaffinization, the next steps involved probe hybridization, GEM generation, and library construction as outlined in the "Chromium Fixed RNA Profiling Reagent Kits for Multiplexed Samples" manual (CG000527; Rev E; 10x Genomics).