Figure 3.

c-Met mediated soluble factors regulate neutrophil mobilization, maturation, and lifespan (A) Neutrophils isolated from mouse bone marrow were treated by indicated CM or RPMI for 48 h. Neutrophils were then stained by CD11b and Ly6G antibodies, followed by flow cytometry analysis. Left, representative flow cytometry graph. Right, quantification of CD11b^hi/Ly6G^hi percentage. (B) Neutrophils were seeded in the upper chamber of a trans-well with indicated 1st CM or RPMI in the lower chamber. Number of neutrophils that migrated to the lower chamber was counted after 3 h. Left, representative picture at the endpoint. Right, quantification of cells migrated to the lower chamber. (C) Neutrophils isolated from mouse bone marrow were treated by CM or RPMI for 48 h. Neutrophils were stained by 7AAD and AnnexinV and followed by flow cytometry analysis. Left, representative flow cytometry graph. Live cells that were not undergoing apoptosis were gated in the lower left panel. Right, quantification of survived neutrophils after 48 h (** p  <  0.01, or *** p < 0.001).