Western Blot Analysis of M. xanthus Proteins

Samples were prepared by harvesting exponentially growing M. xanthus cultures and subsequent resuspension in SDS lysis buffer to an equal concentration of cells. Western blot analyses were performed as described⁶⁵ with rabbit polyclonal α-PomX (1:10000)^{31 (link)}, α-PomY (1:10000)^{31 (link)}, α-PilC (1:3000)^{68 (link)}, or α-mCh (1:10000; Biovision) primary antibodies together with horseradish-conjugated goat α-rabbit immunoglobulin G (1:25000) (Sigma-Aldrich) as the secondary antibody. For PomY blots, protein transfer was performed with a Tris/CAPS buffer system (BioRad). Protein transfer was done with Trans-Blot Turbo buffer (BioRad) for all other blots. Blots were developed using Luminata Forte Western HRP Substrate (Millipore) and visualized using a LAS-4000 luminescent image analyzer (Fujifilm).