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. 2008 Feb 15;190(8):2851–2857. doi: 10.1128/JB.01904-07

FIG. 6.

FIG. 6.

Immunodetection and localization of SprB. (A) Total cell extracts examined for SprB by SDS-PAGE and Western blot analysis. The extract in lane 1 was prepared by boiling cells in SDS-PAGE loading buffer, and the extracts in lanes 2 to 6 were prepared by disruption of cells by passage through a French pressure cell, followed by boiling of the extract in SDS loading buffer. Lanes 1 and 2, wild-type F. johnsoniae FJ1; lane 3, sprB mutant FJ156; lane 4, sprB mutant FJ117; lane 5, FJ156 complemented with pSN60; lane 6, FJ117 complemented with pSN60. The arrow indicates the position of the highest-molecular-weight form of SprB. Fifty micrograms of protein was loaded in each lane. (B) Detection of SprB in cell fractions by Western blot analysis. Lane 1, whole-cell extract of wild-type strain FJ1; lane 2, whole-cell extract of sprB mutant FJ156; lane 3, soluble fraction of FJ1; lane 4, insoluble (membrane and particulate) fraction of FJ1; lane 5, concentrated spent growth medium of FJ1. The arrow indicates the position of the highest-molecular-weight form of SprB. Equal amounts, corresponding to 30 μg of cell protein of starting material, were loaded in the lanes.