Western blot analysis of PMN-derived HGF: effect of NaCl pretreatment.

PMNs were stimulated with cytochalasin B and fMLP at 37°C; the supernatant was then incubated for 30 minutes at 37°C with 2 M NaCl (NaCl⁺) or without NaCl (NaCl⁻) and then subjected to Western blot analysis in nonreducing conditions and revealed with antihuman HGF monoclonal antibodies; rh-HGF and HGF derived from purified specific granules (gr) were loaded and blotted in parallel. Note that both pro-HGF and mature HGF yielded a 92-kd band (arrow) in nonreducing conditions. These data are representative of one experiment of 3.