FIG 6.

Motility analyses of Δeps mutants. (A) Colony-based motility assay of Δeps mutants. T4P-dependent motility and gliding motility were tested on 0.5% and 1.5% agar, respectively. Images were recorded after 24 h. Scale bars, 1 mm, 1 mm, and 10 μm (left to right). (B and C) T4P shear-off assay. Immunoblot detection of the major pilin PilA in sheared T4P (top) and in total cell extract (middle), where the same number of cells grown on 1% CTT, 1.5% agar was loaded per lane. The top and middle blots were probed with anti-PilA antibodies (calculated molecular mass, 23.4 kDa). The middle blot was stripped and probed with antibodies against PilC (calculated molecular mass, 45.2 kDa) as a loading control. (D) Immunoblot detection of proteins of the T4P machinery using anti-PilA, -B, -C, -M, -N, -O, -P, -Q, -T, -Tgl, and -TsaP antibodies. The same number of cells coming from exponentially growing liquid cultures was loaded per lane. As a negative control, cells containing a single in-frame deletion mutation in the relevant gene were used.