Fig. 2

Co-overexpression of BAM improves surface display of HbpD-ELL. A Flow cytometric data showing the fluorescent intensity of cells bound by anti-ESAT6 monoclonal antibodies. The different culture conditions tested (i.e. HbpD-ESAT6 or HbpD-ELL uninduced, induced, or induced for BAM as well) are indicated on the right. B Fold change of surface display of cells expressing HbpD constructs as in (A). Graph shows the mean from three independent flow cytometry experiments. C Schematic representation and passenger models for HbpD-SpT2 and HbpD-SpT2-LL. Coupling of SpT2 to externally added SpC2-mScarlet allows detection of surface displayed HbpD-SpT2 and HbpD-SpT2-LL. D Coomassie-stained SDS-PAGE gels of whole cell lysates from E. coli BL21 (DE3) expressing HbpD-SpT2-LL with or without co-expression of BAM compared to lysates of cells expressing HbpD-SpT2 only. E Coomassie-stained SDS-PAGE gels of whole cell lysates from E. coli BL21 (DE3), expressing HbpD-SpT2 and HbpD-SpT2-LL and incubated with SpC2-mScarlet for 21 h at 4 °C. HbpD variants and SpC2-mScarlet adducts (<), BamA (<) and non-bound SpC2-mScarlet(*) are indicated. Panel E is composed of lanes taken from the same image of a Coomassie-stained SDS-PAGE gel; the assembled parts are boxed