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Loss of O-glycosylation altered the levels of pilin in R. solanacearum. Whole-cell lysates of different R. solanacearum strains were run on SDS–PAGE gel followed by immunoblotting using rabbit polyclonal anti-pilin and mouse monoclonal anti-RNA polymerase (1:2500, RNAP ɑ-subunit; Neoclone). Membranes were then probed with IRDye conjugated anti-mouse and anti-rabbit antibodies and visualized on an Odyssey infrared imaging system (LI-COR Biosciences, Lincoln, NE).
