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. 2005 Jul;25(14):6279–6288. doi: 10.1128/MCB.25.14.6279-6288.2005

FIG. 1.

FIG. 1.

PDGF D overexpression and activation by a protease secreted by LNCaP and PC3 cell lines. (A) PC3-neo, PC3-PDGF D, LNCaP-neo, and LNCaP-PDGF D cells were grown to 100% confluence in complete media, washed once with PBS, and then cultured in serum-free medium for 48 h. CM was collected, and proteins were resolved using SDS-PAGE under reducing conditions. (B) Three milliliters of CM from PC3-neo and PC3-PDGF D was concentrated using Ni-NTA agarose beads and resolved on a reducing SDS-PAGE gel (see Materials and Methods). Anti-PDGF D-growth domain (GD) antibody was used for immunoblot analysis. FL-M, full-length PDGF D protein monomer; GD-M, PDGF D growth domain monomer. (C) Semiquantitative RT-PCR analysis was performed to examine RNA levels of PDGF D in parental cell lines, as well as neo control and PDGF D overexpressing cell lines. The primer pair used detected both endogenously and exogenously expressed PDGF D.