Fructose-1,6-bisphosphatase 1 mediates Ikkβ dephosphorylation and promotes the polarization of immunosuppressive tumor-associated macrophages during metabolic adaptation in gallbladder carcinoma

Human GBC tissues were obtained from patients undergoing surgery at the Department of General Surgery, Shengjing Hospital of China Medical University. The tumor samples were processed for single-cell RNA sequencing and GBC organoid construction. To prepare the tissues for single-cell RNA sequencing, tissues were dissociated into single cells in dissociation solution in a 37°C water bath with shaking for 20 min at 100 rpm. The cell pellet was subsequently resuspended in 50 μl of 1× PBS. Single-cell suspensions were loaded into a 10x Chromium system according to the manufacturer’s instructions of the 10X Genomics Chromium Single-Cell 3’ kit (V3). The following cDNA amplification and library construction steps were performed according to the standard protocol.