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ConA lectin probing following PNGase or mannosidase treatment of released E. coli surface proteins. LF: AIEC LF82, MG: E. coli K-12 MG1655. NT, Non-acid hydrolyzed; AH, Acid hydrolyzed; L, Molecular weight ladder; Mock, Mock-treated sample; Man, Mannosidase-treated sample. (A) PNGase F treatment probed with ConA. (B) Anti-type I fimbria staining of membrane (A) . (C) Mannosidase treatment probed with ConA.

Journal: Frontiers in Microbiology

Article Title: Biochemical characterization of the Escherichia coli surfaceome: a focus on type I fimbriae and flagella

doi: 10.3389/fmicb.2025.1507286

Figure Lengend Snippet: ConA lectin probing following PNGase or mannosidase treatment of released E. coli surface proteins. LF: AIEC LF82, MG: E. coli K-12 MG1655. NT, Non-acid hydrolyzed; AH, Acid hydrolyzed; L, Molecular weight ladder; Mock, Mock-treated sample; Man, Mannosidase-treated sample. (A) PNGase F treatment probed with ConA. (B) Anti-type I fimbria staining of membrane (A) . (C) Mannosidase treatment probed with ConA.

Article Snippet: To further probe the nature of the attached glycans, protein aliquots were subjected to PNGase F or a broad range mannosidase (α1,2/3/6) (New England Biolabs, France) digestion according to the manufacturer’s specifications, with the exception that incubations were carried out overnight for 18 h. Mock treatments were identical, maintaining the initial denaturation step, but substituting water for enzyme.

Techniques: Molecular Weight, Staining, Membrane

Anti-type I fimbria staining following PNGase F or mannosidase treatment of released E. coli surface proteins. PNGase F or mannosidase treatment of released E. coli surface proteins increased anti-type I pili detection. L: PageRule protein ladder; LF: AIEC LF82; MG: E. coli K-12 MG1655; − denotes mock-treated, + denotes enzyme treated, C denotes enzyme-only control.

Journal: Frontiers in Microbiology

Article Title: Biochemical characterization of the Escherichia coli surfaceome: a focus on type I fimbriae and flagella

doi: 10.3389/fmicb.2025.1507286

Figure Lengend Snippet: Anti-type I fimbria staining following PNGase F or mannosidase treatment of released E. coli surface proteins. PNGase F or mannosidase treatment of released E. coli surface proteins increased anti-type I pili detection. L: PageRule protein ladder; LF: AIEC LF82; MG: E. coli K-12 MG1655; − denotes mock-treated, + denotes enzyme treated, C denotes enzyme-only control.

Article Snippet: To further probe the nature of the attached glycans, protein aliquots were subjected to PNGase F or a broad range mannosidase (α1,2/3/6) (New England Biolabs, France) digestion according to the manufacturer’s specifications, with the exception that incubations were carried out overnight for 18 h. Mock treatments were identical, maintaining the initial denaturation step, but substituting water for enzyme.

Techniques: Staining, Control