Fig. 2.

IL-25 activates alternative macrophages (M2), which promote HCC cell migration and invasion. a IL-25 was used to directly treated HCC cells in vitro, the relative value of CCK-8, Annexin-V, Brdu, Transwell experiment. (Please see results in Additional file 1: Figure S1A-D). b Immunofluorescent staining was performed in normal liver tissues (n = 55) and HCC tumor tissues (n = 98). Representative immunofluorescence images. Bar, 50 μm. c Immunohistochemistry staining was performed in a HCC tissue microarray (n = 98). Correlation between M2 macrophage (CD206/CD68) percentage and IL-25 scores. CD206 is an M2 macrophage marker, and CD68 is macrophage marker. d and e Macrophages (derived from THP-1) were treated with IL-25 in a time- and concentration-dependent manner. Tumor necrosis factor-α (TNF-α) and CD206 were examined by Western blotting. f and i HCC cells co-cultured with M0 or M2 macrophages. f Cell growth was determined with the Brdu kit. Statistical data were shown at the right. Bar, 100 μm. HCC cells migration (g) and invasion (h) were determined by Transwell assay. Statistical data were shown at the right. Bar, 100 μm. i Mesenchymal maker vimentin, EMT regulator Snail, epithelial marker E-cadherin, extracellular signal-regulated kinase (ERK), and p-ERK were detected by Western blotting. ***p < 0.001, ns, no significance