Figure 8

Activation of the Wnt signaling pathway by macrophages recruited to the liver by inflammation. (a) Flow cytometry analysis to determine the proportions of resident macrophages (Kupffer cells) and macrophages recruited to the liver of TAA-administered mice. Kupffer cells were isolated as F4/80^high and CD11b^low cells, and recruited macrophages as F4/80^low and CD11b^high cells. (b) The percentages of Kupffer cells and recruited macrophages in the liver of control mice and TAA-administered mice (left and middle). Relative expression of Wnt3a derived from recruited macrophages in the liver of control mice and TAA-administered mice (right). (c) Scheme of the LPS stimulation assay (left). U937 human macrophage cells were activated by treatment with 5 μg/ml LPS for 48 h. U-937 cells treated with PBS were used as a negative control. Activated macrophage conditioned medium (AMCM) and non-activated macrophage conditioned medium (NAMCM) were collected. The NCC-CC1 IHCC cells and IHCC organoids were then treated with AMCM or NAMCM for 48 hours. Relative expression of WNT3A in the U937 human macrophage cell line untreated or treated with LPS was examined (right). (d) Western blot analysis of nuclear and cytosolic β-catenin (CTNNB1) in NCC-CC1 cells (left) and IHCC organoids (right) treated with NAMCM or AMCM. β-Actin (ACTB) was used as an internal control.