FIGURE 4.

CG regulates the secretion of full-length CRAMP from mouse neutrophils. A, loss of cell-associated CRAMP during neutrophil activation. TNF-α-primed, IC-stimulated WT, and CG/NE neutrophils were harvested at the indicated time points, and cell-associated CRAMP was analyzed by Western blot (WB) using a C terminus-directed goat antibody. B, hCG cleaved recombinant full-length murine CRAMP (SUMO FL-CRAMP) to release C-terminal peptides as detected with a C terminus directed anti-c-Myc antibody. C, CRAMP release into the air pouch was dependent on CG. Total proteins from acetone-precipitated supernatants obtained as described above were probed for CRAMP by Western blot using an antibody directed against the C terminus (C-term). The CRAMP doublet of 15 and 20 kDa has been described in human neutrophils and seminal plasma (47, 54). NS denotes a nonspecific band.