Fig. 2
LBP promotes GC-LM in vivo. A Protein level of LBP was determined in the cell lysate and CM of GC cell lines and normal gastric epithelial cell line (GES-1) by WB. Ponc. S staining was used as the loading control of CM. CM, conditioned medium. Ponc. S, Ponceau S staining. B Representative IF images of LBP expression in GES-1 and GC cells (AGS, HGC-27, MKN45). C Schematic diagram of intrasplenic injection mouse model of LM and experimental workflow. The image was drawn by using BioRender.com. D Representative bioluminescent images (left) and BLI quantification (right) of mice with intrasplenic injection of AGS cells (1 × 106 cells per mouse) with or without LBP stable overexpression for LM analysis (n = 5 mice per group for control and overexpression). E Mice were euthanized in the 5th week after intrasplenic injection. Representative bioluminescent images (left) and photon flux quantification (right) of LM are shown. F Representative photographs (left) and H&E staining with enlargement (middle, right) of LM are shown. G-K Intrasplenic injection of MKN45 cells (1 × 10.6 cells per mouse) with or without LBP stable knockdown for LM analysis (n = 6 mice per group). Representative bioluminescent images (G, left) and BLI analysis (G, right) of mice are shown. Mice were euthanized in the 4th week after intrasplenic injection. Representative bioluminescent images (H, column of IVIS image), photographs (H, column of whole liver) and H&E staining with enlargement (H, columns of H&E and enlargement) of LM are shown, respectively. Quantification of liver photon flux (I), liver weight (J) and surface metastatic nodules of liver (K) was applied. Scale bars are 20 μm (B), 1.0 cm (F left, H column 2), 3 mm (F middle, H column 3), 300 μm (F right, H column 4), and the colored scale bars represent the intensity of luminescence as indicated, respectively. Data are representative of three independent experiments. Data are shown as mean ± SEM (D, G), or mean ± SD (E, I, J, K). p values were determined by two-way ANOVA test (D, G), two-tailed unpaired Student’s t test (E), or one-way ANOVA test (I, J, K) (*P < 0.05, **P < 0.01, ***P < 0.001)