Fig. 2

Expression and surface display of the full-length lipoproteins in E. coli, at 37 °C. a Western blot of whole-cell lysates showing full-length lipoproteins. Lane 1: Marker, Lane 2: NmCsgG, Lane 3: NmMtrC, Lane 4: NmBamE, Lane 5: Nm putative lipoprotein, Lane 6: EcBamE, Lane 7: EcLolB, Lane 8: EcLptE and Lane 9: EcPal. FLAG-tag specific antibodies were used for detection. b, c FACS analysis of E. coli expressing full-length lipoproteins at 37 °C. At 37 °C, E. coli BL21DE3 (pET15b) expressing N. meningitidis lipoproteins: CsgG, MtrC, BamE and a putative lipoprotein (b) and E. coli lipoproteins (c): BamE, LolB, LptE and Pal were incubated with the monoclonal anti-FLAG antibody. The grey areas represent the fluorescence signals obtained with the control (BL21DE3-pET15b ∅). The coloured lines represent the full-length lipoproteins. Panel B (heterologous lipoproteins): Purple: CsgG, Dark Red: MtrC, Light Blue: nmBamE, Dark Green: putative lipoprotein; Panel C (homologous lipoproteins): Dark Blue: BamE, Light Green: LolB, Orange: LptE, Light Red: Pal