Figure 2: CXCR4 promotes hepatoma cell proliferation and survival in vitro.

(a) Western blot analysis of CXCR4 and GAPDH for Huh7, PLC/PRF/5, Hep3B, HepG2, SNU448 and SK-Hep1 cells (upper panel), for Huh7 cells stably transfected with empty vector or CXCR4 (lower left panel) and for SK-Hep1 cells stably transfected with nonspecific shRNA (shCon) or CXCR4-specific shRNA (shCXCR4) (lower right panel). Data are representative immunoblots of three independent assays. (b) Cell proliferation analysis for Huh7 cells without or with stably CXCR4 overexpression, SK-Hep1 cells without or with stably CXCR4 knockdown, SK-Hep1 cells without or with AMD3100 (100 nM) or CXCR4-neutralization antibody (100 μg ml⁻¹) treatment (n=3). Student’s t-test, *P<0.05. Error bars in panels are defined as s.d. (c) Representative micrographs and quantification of soft agar colonies for above-mentioned hepatoma cells (n=3). Student’s t-test, *P<0.05. Error bars in panels are defined as s.d. (d) Representative micrographs and quantification of the above-mentioned hepatoma cells in the Transwell migration assay (n=3). Student’s t-test, *P<0.05. Error bars in panels are defined as s.d. (e) Representative dot plots of flow cytometric analysis of above-mentioned hepatoma cells cultured in serum-deprivation medium for 48 h. Cells were subjected to annexin-V and Propidium lodide (PI) staining, and the percentage of cells displaying annexin-V single positive and annexin-V, PI double positive are denoted (n=3). Student’s t-test, *P<0.05. Error bars in panels are defined as s.d.