Fig. 5

GAL-8 and CD73 interference restraints the function of circHMGCS1–016 in ICC cells. A. The relative level of CD73 and GAL-8 was measured by qRT-PCR in the ICC cells; Data are representative of 3 independent tests; B. The expression of CD73 and GAL-8 were modified by lentivirus-mediated knockdown in RBE-circHMGCS1–016 cells. Data are representative of 3 independent tests(*** p < 0.001); C. The efficacy of GAL-8/CD73 interference was analyzed by western blot; D. Invasion assay was performed to detect the invasion ability of RBE-circHMGCS1–016 cells and RBE-circHMGCS1–016 cells with CD73 and GAL-8 interference (Bar = 200 μm). Data are representative of 3 independent tests(*** p < 0.001); E. Colony formation assay was performed to detect the ability of colony formation in RBE-circHMGCS1–016 cells and RBE-circHMGCS1–016 cells with CD73 and GAL-8 interference. Data are representative of 3 independent tests(*** p < 0.001); F. The level of GAL-8 in the supernatant of RBE-circHMGCS1–016 cells and RBE-circHMGCS1–016 cells with CD73 and GAL-8 interference; Data are representative of 3 independent tests(*** p < 0.001); G. The level of adenosine concentration in the supernatant of RBE-circHMGCS1_016 cells and RBE-circHMGCS1–016 cells with CD73 and GAL-8 interference; Data are representative of 3 independent tests(** p < 0.01); H. A co-culture showed the supernatant from RBE-circHMGCS1–016 cells inhibited the CD4⁺ and CD8⁺ T cell proliferation compared to RBE-circHMGCS1–016 cells with CD73 and GAL-8 interference (*** p < 0.001); I. Chemokine chips and ELISA were employed to determine the different chemokines in the supernatant between RBE-circHMGCS1–016 cells and RBE-circHMGCS1–016 cells with CD73 and GAL-8 interference; J. A model for circHMGCS1–016 driven ICC development and established the immune privilege microenvironment