Figure 2

Changes in CXCR4 and Met expression after HGF treatment implicated HDACs. (A) The cells were transiently transfected with pCXCR4(−2632/+86)Luc. Some starved cells, treated with 2.5 μ M TSA, were added with HGF (200 ng ml⁻¹) for 1 day. The other cells in 10% serum were treated with 2.5 μ M TSA for 1 or 2 days. The histograms indicate the absolute values for ratios of Firefly/Renilla luciferase activity. The data are the means±s.e. of three independent experiments performed in triplicate. ^**P<0.005 and ^***P<0.001 vs respective control value; ^ΔP<0.05 vs HGF-treated MDA-MB231 cells. (B) Western blot analyses of CXCR4 protein levels in starved (0.1% serum) and serum-maintained (10% serum) cells treated with 2.5 μ M TSA for 1 day. The numbers at the bottom indicate the fold variations relative to the value of starved (st) MCF-7 cells. Immunoblot with anti-vinculin antibody was used for normalisation. The experiments were repeated three times, with similar results. (C) Western blot analyses of CXCR4 protein levels in cells treated with HGF with or without 2.5 μ M TSA for 1 or 2 days (d). Representative western blots are shown, and the numbers at the bottom indicate the fold variations relative to the value of st MCF-7 cells. Immunoblot with anti-vinculin antibody was used for normalisation. The experiments were repeated five times, with similar results, and the means±s.e. are reported in the histograms. ^*P<0.05, ^**P<0.005 and ^***P<0.001 vs respective control values; ^ΔΔP<0.005 vs HGF-treated cells. (D) Western blot analyses of Met protein levels in cells treated with HGF with or without 2.5 μ M TSA for 1 or 2 days (d). The numbers at the bottom indicate the fold variations relative to the respective st values. Immunoblot with anti-vinculin antibody was used for normalisation. The blots are representative of experiments repeated three times.