HCC tissues were mixed with 0.4µl of lysis buffer and incubated on ice for 10 min. After triturating the tissue samples, tissue lysates were centrifuged at 500 rcf for 5 min at 4°C. Emerging supernatant was removed followed by resuspending the tissue lysates in 1 ml nuclei wash and resuspension buffer. Subsequent to the evaluation of cell viability, the single cell suspension underwent the process of cell nuclei sorting using Fluorescence-Activated Cell Sorting (FACS). For cDNA synthesis and preparation of single-nucleus RNA seq libraries, Single nucleus suspensions were processed using the Chromium Next GEM Single Cell 3' v3.1 (Dual Index) kit to achieve single-cell resolution within a droplet-based system. Following sequencing steps such - selection of barcoded single cells, reverse transcription and PCR was performed subsequently.  Agilent Bioanalyzer was employed to ensure sample quality to establish a sequencing library effectively. Construction of sequencing libraries was conducted on Illumina NovaSeq platform. snRNA-seq