Journal: PLOS Biology
Article Title: Structure of human glycoprotein 2 reveals mechanisms underlying filament formation and adaption to proteolytic environment in the digestive tract
doi: 10.1371/journal.pbio.3003238
Figure Lengend Snippet: a , Western blotting analysis of GP2 derived from human pancreas (donor 1, before and after elastase cleavage) and small intestines (donors 2–4, without elastase treatment), probed with two GP2 antibodies, anti-GP2 (35–179) and anti-GP2 (111–387). −E, GP2 incubated without elastase; +E, GP2 incubated with elastase; +E’, the same sample as +E, but the loading volume for SDS–PAGE was reduced to one-fifth; Mw, molecular weight. b , Representative negative-stain EM images of (e) GP2 derived from the pancreas. Scale bar represents 50 nm. c , The SDS–PAGE of interaction assays between FITC-FimH L and GP2. The bands represent FITC-FimH L that were visualized with fluorescence. Samples were either washed using centrifugal filters (+ in the row labeled “filter”) or directly analyzed using SDS–PAGE without wash (− in the same row) as controls. d , Representative negative-stain EM images of (e) GP2 incubated with or without FimH L , labeled with immunogold and FimH antibodies. White arrows label GP2 filaments and the black arrow labels immunogold beads. Scale bar represents 50 nm. e , Representative light microscopy images of (e) GP2-mediated E. coli cell aggregation. Scale bar represents 10 μm. All assays were repeated independently at least three times with similar results. The data underlying this figure can be found in .
Article Snippet: E. coli TOP10 competent cells (Yeasen, catalog number 11801ES80) were used in this study for GP2-induced aggregation assay.
Techniques: Western Blot, Derivative Assay, Incubation, SDS Page, Molecular Weight, Staining, Fluorescence, Labeling, Light Microscopy