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. 2013 Sep 24;21(12):2195–2204. doi: 10.1038/mt.2013.191

Figure 3.

Figure 3

CGKRK binding to candidate receptor proteins. (a) Binding of FAM–CGKRK to purified p32, NLC3, CKAP4, and nardilysin1 (NRD1). Binding of increasing amounts of FAM–CGKRK peptide and a control FAM–CREKA peptide to immobilized p32, NLC3, CKAP4, and NRD1 proteins was detected by fluorescence and normalized to nonspecific binding to the plastic plate. One representative binding curve out of three independent experiments with similar results is shown. The Kd was calculated from all three experiments using Prism software. (b) Binding and internalization of CGKRK phage particles in a series of MCF10 human breast tumor cells. MCF10 cells were incubated with phage displaying CGKRK or a polyglycine control peptide, CG7C, overnight at 4 °C or for 1 hour at 37 °C. To assess internalization, phages bound at the cell surface were removed by washing the cells with an acid buffer before phage titration. Statistical analyses were performed with Student's t-test. n = 3; error bars, mean ± SEM; **P < 0.01. (c) Total expression levels of NRD-1, NLC3, CKAP4, and p32 in the MCF10 cells assessed by immunoblotting.