Fig. 1

A-HSCs expressed high levels of IL-8. A Immunofluorescent staining of primary human a-HSCs isolated from a representative sample of HCC with an anti-α-SMA antibody,anti-vimentin antibody, and IgG. Scale bar, 50 μM. B The levels of various angiogenic chemokines in the cell-free culture supernatants of the a-HSCs were measured by Multiplex bead–based enzyme–linked immunosorbent assay at day 2. The data are expressed as mean ± SEM of triplicates. (C) The concentrations of IL-8 (pg/mL) in the supernatants of a-HSCs and hepatoma cells were determined by ELISA. The concentration of IL-8 in the a-HSCs culture medium was markedly higher compared with hepatoma cells. (D) The expression of IL-8 in the abovementioned cells was assessed by Western blotting. A-HSCs spontaneously released large amounts of IL-8, whereas hepatoma cells produced low levels of IL-8. (E) Hepatocarcinoma samples were stained with various antibodies, and different levels of antibodies expression can be seen on the same section. E (a) Immunohistochemical staining for α-SMA (1:1000) to identify a-HSCs in the tumour stromal region. E (b) The IL-8 distribution in hepatocarcinoma samples was visualized by immunohistochemical staining using the IL-8 neutralizing antibody (1:400). E (c) Neovascularization in hepatocarcinoma samples was visualized by immunohistochemical staining for CD31 (1:400). Scale bar 200 μM. One of the 22 representative micrographs is shown