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.人类胆囊癌（Gallbladder Carcinoma, GBC）组织采集自中国医科大学附属盛京医院普通外科接受手术治疗的患者。上述肿瘤样本将用于单细胞RNA测序以及GBC类器官构建。为制备用于单细胞RNA测序的组织样本，将组织置于解离液中，于37℃水浴环境下以100 rpm转速振摇20分钟，解离为单个细胞。随后将细胞沉淀重悬于50 μl的1×磷酸盐缓冲液（Phosphate Buffered Saline, PBS）中。按照10X Genomics公司Chromium单细胞3'测序试剂盒（V3版）的制造商说明书，将单细胞悬液上样至10x Chromium系统中。后续的cDNA扩增与文库构建步骤均按照标准实验流程完成。