Fig. 4

LBP promotes GC cell colonization and outgrowth in the liver at the early stages by inducing intrahepatic fibrotic PMN formation in pre-modelling mouse model. A Schematic diagram of pre-modelling mouse model of LM and experimental workflow. The image was drawn by using BioRender.com. B Representative bioluminescent images (left) and BLI analysis (right) of mice with intrasplenic injection of AGS cells (1 × 10⁶ cells per mouse) after mice were pretreated with reLBP or IgG for two weeks (n = 5 mice per group). C-G Colonization and proliferation of cancer cells in the liver at the early stage (in the first week) with intrasplenic injection of HGC-27 cells after mice were pretreated with PBS, IgG or reLBP, respectively (n = 10 mice per group). BLI quantification (C) are shown by comparing the bioluminescence at sequential timepoints. Mice were euthanized on the 7th day after intrasplenic injection. Representative photograph and H&E staining (D), IF analyses of GFP⁺ HGC-27 cells in the liver sections (E), flow cytometry analysis of GFP.⁺ HGC-27 cells in the liver (F), and representative IHC images of Ki67 in tumour cells in the liver are shown. H Representative H&E and IF images of myeloid cells (Ly6G, F4/80), α-SMA, FN, and COL1 in the livers after mice were pretreated with PBS, IgG or reLBP for two weeks, respectively. Scale bars are 1.0 cm (D, left), 200 μm (H, H&E), 100 μm (D-right, E, H-IF) and 50 μm (G), the colored scale bars represent the intensity of luminescence as indicated, respectively. Data are representative of three independent experiments. The BLI quantification is shown as mean ± SEM at sequential timepoints (B, C). Data of flow cytometry analysis are shown as mean ± SD (F). p values were determined by two-way ANOVA test (B, C) or one-way ANOVA test (F) (ns, not significant, * P < 0.05, ** P < 0.01, *** P < 0.001)