Figure 1.
Assembly of PulG pili in mutants lacking single minor pseudopilins. (A) PulG immunodetection in 0.005 OD600 nm units of cell and sheared fractions (C, SF) of E. coli carrying the pulG gene in a plasmid (pulG alone), or the pul genes (WT) on plasmid pCHAP231 or its minor pseudopilin gene deletion derivatives. LamB porin is used as a lysis control. (B) PulG percentage in SF (mean+s.d.) from three independent experiments like the one in (A). NA, not applicable. (C) Immunofluorescence (IF) and phase contrast microscopy of strains analysed in (A). PulG staining is in green and DAPI in magenta. Scale bar, 5 μm. (D) Box plot of normalized number of pili observed in (C) from ∼40 randomly selected areas in two independent experiments. (E) PulA immunodetection in cell extracts and supernatants (C, SN) of bacteria carrying pulA on a plasmid (+pulA), or expressing all pul genes on plasmid pCHAP8185 (WT) or its minor pseudopilin single deletion derivatives. (F) Percentage of secreted PulA (mean+s.d.) from five independent experiments like the one in (E). Figure source data can be found in Supplementary data.