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. 2004 Mar;186(6):1658–1666. doi: 10.1128/JB.186.6.1658-1666.2004

FIG. 6.

FIG. 6.

Detection of mixed DotB/DotB K162Q multimers. Lysate from L. pneumophila Flag-DotB strain JV3079 containing one of three plasmids was subjected to Ni-NTA-agarose. Strains were JV3079 with empty vector pJB908 (lanes 1 to 3), JV3079 with the His-DotB complementing clone pJB1192 (lanes 4 to 6), and JV3079 with the His-DotB K162Q clone pJB2444 (lanes 7 to 9). For each strain, three samples were taken: total protein (lanes 1, 4, and 7), Ni-NTA-agarose flowthrough (lanes 2, 5, and 8), and Ni-NTA-agarose eluate (lanes 3, 6, and 9). Samples were separated via SDS-PAGE, blotted onto a polyvinylidene fluoride membrane, and used in a Flag Western blot to detect the presence of Flag-DotB. The molecular masses of relevant markers (in kilodaltons) are on the left.