Immunoblotting was performed as described previously (35 ). For sample preparation, M. xanthus cells were harvested from exponentially growing suspension cultures and resuspended in SDS lysis buffer. Proteins were loaded from an equal number of cells per sample. As primary antibodies, rabbit polyclonal anti-FLAG (1:2,000; Rockland) and anti-PilC (1:2,000) (38 (link)) antibodies were used, with horseradish peroxidase (HRP)-conjugated anti-rabbit immunoglobulin G (1:15,000, Sigma) as the secondary antibody. Immunoblots were developed using Immobilon Forte Western HRP substrate (Millipore) on a LAS-4000 imager (Fujifilm).
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