Lu/BCAM Adhesion Glycoprotein Is a Receptor for Escherichia coli Cytotoxic Necrotizing Factor 1 (CNF1) (original) (raw)
Figure 4
CNF1 directly interacts with Lu/BCAM on the cell surface.
Suspensions of HeLa cells ((A–C) 1×105 cells in 1 ml medium) were incubated for 20 min at 4°C with 2 µg DyLight488-labeled GST-CNF1 (CNF1DL488) alone or together with increasing amounts of rBCAM protein (A). Cells were then washed in PBS and subjected to FACS analysis. The experiment was repeated with 2 µg DyLight488-labeled GST-CNF1 (CNF1DL488) alone or together with increasing amounts of αLu/BCAM antibody (molar ratio CNF∶antibody up to 1∶50, * p<0.05) (B) or a 50-fold excess of the αLRP antibody as indicated (C). Data are presented as % fluorescence of control with CNF1DL488 set to 100%. Non- (D) and Lu/BCAM-expressing (E) K562 cells were either left untreated (mock) or were incubated with 2 µg CNF1DL488 for 20 min at 4°C and, following washing with PBS, subjected to FACS analysis. Results are presented as histogram plots, where single cell events are plotted against cell surface-bound fluorescence (Log FL intensity).