Figure 1
HGF and c-Met are up-regulated by G-CSF in a CXCR4/SDF-1–dependent manner. (A) Plasma HGF levels, evaluated by ELISA, in mice that received 1 injection of G-CSF and were killed 24 hours afterward or in mice receiving 5 consecutive injections of G-CSF. (B) Mice were given G-CSF for 5 consecutive days. HGF intracellular levels were determined in BM leukocytes by FACS analysis. (C) Intracellular HGF levels in BM PMN cells of control mice or mice receiving 3 or 5 consecutive injections of G-CSF were determined by FACS analysis. (D) G-CSF knockout (KO) mice or wild-type (WT) counterparts were injected with PBS or with 3 or 5 consecutive daily injections of G-CSF. The levels of circulating SKL cells were determined. (E) Plasma HGF levels in G-CSF KO and WT mice receiving PBS or 5 consecutive injections of G-CSF were determined by ELISA. (F) Cell-surface c-Met levels were quantified by FACS analysis. Representative c-Met expression on total BM cells of control (b) or 5 days of G-CSF in treated mice (c), together with control secondary staining (a) is shown. (G) A summary of 4 independent experiments showing membrane c-Met expression on BM cells, PMN cells, BM c-kit+/Lin− cells, or SKL/CD34− cells. (H-I) BM cells were left untreated or treated in vitro with SDF-1. c-Met activation (H) and intracellular HGF expression (I) were measured. (J) Circulating SKL cells in control mice or mice receiving 3 or 5 injections of G-CSF alone, or together with anti CXCR4 neutralizing Abs on the last 2 days of G-CSF treatment. (K) Percentage of phospho–c-Met on SKL cells in the PB of control mice, mice treated with 5 injections of G-CSF alone, or with anti-CXCR4 neutralizing Abs on days 4 and 5 of G-CSF treatment. (L) Representative FACS analysis showing the percentage of phospho–c-Met expression on circulating SKL cells. Percentages indicate the frequency of phospho c-Met–positive cells. (M) HGF levels in BM PMN cells from control mice or mice receiving 3 injections of G-CSF alone, or with anti-CXCR4 neutralizing Abs on days 2 and 3 of G-CSF treatment. Indicated values are presented as mean ± SE in ≥ 3 independent experiments, 6 mice in each group. *P < .05; **P < .001. SSC indicates side scatter.

HGF and c-Met are up-regulated by G-CSF in a CXCR4/SDF-1–dependent manner. (A) Plasma HGF levels, evaluated by ELISA, in mice that received 1 injection of G-CSF and were killed 24 hours afterward or in mice receiving 5 consecutive injections of G-CSF. (B) Mice were given G-CSF for 5 consecutive days. HGF intracellular levels were determined in BM leukocytes by FACS analysis. (C) Intracellular HGF levels in BM PMN cells of control mice or mice receiving 3 or 5 consecutive injections of G-CSF were determined by FACS analysis. (D) G-CSF knockout (KO) mice or wild-type (WT) counterparts were injected with PBS or with 3 or 5 consecutive daily injections of G-CSF. The levels of circulating SKL cells were determined. (E) Plasma HGF levels in G-CSF KO and WT mice receiving PBS or 5 consecutive injections of G-CSF were determined by ELISA. (F) Cell-surface c-Met levels were quantified by FACS analysis. Representative c-Met expression on total BM cells of control (b) or 5 days of G-CSF in treated mice (c), together with control secondary staining (a) is shown. (G) A summary of 4 independent experiments showing membrane c-Met expression on BM cells, PMN cells, BM c-kit+/Lin− cells, or SKL/CD34− cells. (H-I) BM cells were left untreated or treated in vitro with SDF-1. c-Met activation (H) and intracellular HGF expression (I) were measured. (J) Circulating SKL cells in control mice or mice receiving 3 or 5 injections of G-CSF alone, or together with anti CXCR4 neutralizing Abs on the last 2 days of G-CSF treatment. (K) Percentage of phospho–c-Met on SKL cells in the PB of control mice, mice treated with 5 injections of G-CSF alone, or with anti-CXCR4 neutralizing Abs on days 4 and 5 of G-CSF treatment. (L) Representative FACS analysis showing the percentage of phospho–c-Met expression on circulating SKL cells. Percentages indicate the frequency of phospho c-Met–positive cells. (M) HGF levels in BM PMN cells from control mice or mice receiving 3 injections of G-CSF alone, or with anti-CXCR4 neutralizing Abs on days 2 and 3 of G-CSF treatment. Indicated values are presented as mean ± SE in ≥ 3 independent experiments, 6 mice in each group. *P < .05; **P < .001. SSC indicates side scatter.

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