Click on image to view larger version.
FIGURE 2. (A) Amino acid sequence of peptide FROP-1 isolated by peptide phage display. (B) In vitro binding assay with FROP-1. FRO82-2 and MCF-7 cells were grown for 24 h. 125I-FROP-1 was added to wells and incubated for 1 h; as competitor, either 1 x 10–4 M unlabeled FROP-1 or octreotide (Oct) was added immediately before incubation with labeled FROP-1. (C) Internalization of 125I-FROP-1 in MCF-7 cells. Cells were incubated with 1–2 x 106 cpm radioligand for 10 min and 1 h at 37°C. After being washed with acidic glycine buffer (pH 2.8), cells were lysed and internalized radioactivity was measured. Experiments were performed in triplicate; SDs are shown. 1W = first acidic wash, 2W = second acidic wash, L = lysate, C = control with unlabeled peptide; Comp. = competitor.
