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. 2016 Sep 28;113(41):E6228–E6237. doi: 10.1073/pnas.1611594113

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Fig. 6. — Lipoproteins are overexpressed and surface-displayed in LOS-deficient A. baumannii. Intact whole cells (A) or cell lysates (B) were incubated with sulfo-NHS-LC-LC-biotin to biotinylate accessible proteins. After labeling, proteins from the soluble (S) and membrane (M) fractions were subjected to SDS/PAGE and Western blotted using streptavidin-HRP. Intact whole cells containing chromosomal His₆-tag fusions including (C) HMPREF0010_1944 (1944-His₆) (11.52 kDa), (D) HMPREF0010_1945 (1945-His₆) (11.83 kDa), and (E) HMPREF0010_2739 (2739-His₆) (14.76 kDa) were treated with sulfo-NHS-LC-LC-biotin, and proteins were separated using SDS/PAGE. Wild-type, LOS-deficient for each respective mutant, and overexpression strains were blotted using a streptavidin-HRP conjugate (Top), anti-his antibody (Middle), or a polyclonal NADH chain L antibody (Bottom).