Fig. 5

Expression and localization of DDR1 in prostate epithelial cell lines. A Human prostate epithelial cell lines were lysed and resolved by reducing SDS-PAGE followed by immunoblot analyses with anti-DDR1 Sc-532 antibody. CTF: C-terminal fragment of DDR1. B PC-3 malignant and RWPE-1 non-malignant prostate epithelial cells were treated for 90 min with collagen type I, as described in Materials and Methods. The cells were then processed for subcellular fractionation to obtain the cytoplasmic/membrane (B, left panel) and the nuclear (B, right panel) fractions, which were then resolved by reducing SDS-PAGE followed by immunoblot analyses with antibodies D1G6 and E1N8F against total DDR1 and phosphorylated DDR1 (pDDR1), respectively, as described in the Methods section. The blots containing the cytoplasmic/membrane fraction were reprobed with Abs to GAPDH (cytoplasmic) and transferrin receptor1 (TfR1, membrane anchored) whereas the nuclear fraction blots were reprobed with Abs to histone, as controls