Fig. 1: The presence of complex glycans on target cells enhances the binding of Pl-TcdA1. | Nature Communications
a Time course confocal imaging demonstrates that wild type HEK 293T cells bind Pl-TcdA1 (AlexaFluor488 labeled, green) much more readily than their glycosylation-deficient HEK 293 GnTI− counterparts and PNGase F-treated HEK 293T cells. Images were taken after 0, 4, or 8 h of incubation at 37 °C. Scale bar, 100 μm; fluorescence channels are equally thresholded. b Quantification of AlexaFluor488 labeled Pl-TcdA1 binding to wild type HEK 293T (orange squares), PNGase F-treated HEK 293T (blue diamonds), and HEK 293 GnTI− cells (green circles) based on time course confocal microscopy data. Three biological replicates for each cell type were analyzed; error bars represent standard deviation. c Flow cytometry of HEK 293T exposed to AlexaFluor488 labeled Pl-TcdA1 after deglycosylation of cell surface by PNGase F. Histograms of PNGase F-treated and untreated cells, both with and without 100 nM Pl-TcdA1 are shown in comparison. The source data underlying panels (b) and (c) are provided as a Source Data file.