Fig. 3: The identified LptDE-bound lipoproteins are exposed on the surface of E. coli.
From: Surface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteria
A Immunofluorescence assays showing that LptM, Lpp, YedD, YbjP, YajG, Slp, DolP and SlyB are surface-exposed. DAPI and Alexa Fluor 546-conjugated secondary antibody are blue- and red- fluorescencing, respectively. BamB was selected as a negative control. Membrane permeabilization was carried out by treatment with 0.5% Triton X-100 and 5 mM EDTA. Data are representative of three independent experiments. B Dot blot assays (both using intact cells and cell lysates) showing that the identified LptDE-bound lipoproteins (LptM, Lpp, YedD, YbjP, YajG, Slp, DolP and SlyB) are surface-exposed at least for certain percentage of the expressed lipoproteins in the cells. Note that only a small amount of Blc was detected in the lysate, consistent with the low expression levels shown in Fig. 1C. BamB was selected as a negative control, and RcsF was selected as a positive control. With the exception of RcsF, which carries an N-terminal Flag tag, the other lipoproteins are tagged with a C-terminal His tag for visualization or detection. Data are representative of three independent experiments.
