Figure 6
Effect of FliH and FliI deletion on FliC leakage during filament assembly. Measurements of FliC monomers leaked out into the culture media. CBB-staining SDS gels of total extracellular FliC (indicated as T), polymerized FliC (indicated as A), and FliC leaked into the culture media (indicated as S) of (a) SJW1103 (WT), (b) MM1103gK (∆flgK), (c) MMHI0117 (∆fliHI flhB*), (d) MMHI0117-1 [∆fliHI flhB* flhA(F459A)], (e) MMB017 (flhB*) and (f) MMA459 [indicated as flhA(F459A)]. CBB-stained gels were cropped from original image data shown in Fig. S6 in the Supplemental information. The position of 50 kDa molecular mass marker is indicated on the left. (g) Effect of FliH and FliI deletion on assembly of the hook-filament junction at the hook tip. Membrane localization of the MS ring protein, FliF, the transmembrane export gate protein, FlhA, FlgE, and one of the hook-filament junction proteins, FlgL. The membrane fractions of SJW1103, NH004 (∆fliHI flhB* ∆flhA), MMHI0117, or MMHI0117-1 were prepared after sonication and ultracentrifugation. Then, the membrane fractions were subjected to SDS-PAGE and were analyzed by immunoblotting with polyclonal anti-FliF, anti-FlhA, anti-FlgE and anti-FlgL antibodies. Positions of FliF, FlhA, FlgE and FlgL are indicated by arrows. Each immunoblot was cropped from its original image data shown in Fig. S7 in the Supplemental information.
