FIG 2.

Confirmation of PelA and PslG protein production. (A) Total cell extracts (TCEs) from E. coli DH5α/pMJT-pslG_V5 and DH5α/pMJT-pelA_V5 cells grown planktonically to exponential phase. Then, pslG and pelA gene expression was induced upon addition of 1% arabinose for 2 h. A representative image of immunoblot (IB) probed for the presence of V5-tagged PelA or PslG using anti-V5 antibodies is shown. (B) TCEs were used as a loading control. TCE, image of Coomassie-stained SDS-gel after immunoblotting. A total of 20 μg of TCE was loaded. The TCEs from PAO1/pMJT-pslG_V5, PAO1/pMJT-pelA_V5, and PAO1/pMJT-1 strains grown as biofilms for 5 days prior to and after the addition of 1% arabinose for 1 h are shown. Representative images of immunoblot (IB) and Coomassie-stained SDS-gel after immunoblotting (TCE) are shown. Arrows point at a band suggestive of PelA. A total of 20 μg of TCE was loaded. (C) Representative images of immunoblots demonstrating the presence of PslG and PelA in culture supernatants of PAO1/pMJT-pslG_V5 and PAO1/pMJT-pelA_V5 strains grown planktonically to the exponential phase, respectively. Culture supernatants were obtained 1 h after the addition of arabinose. Culture supernatants by PAO1/pMJT-1 were used as a control. The arrow points at a band suggestive of PelA. A total of 10 μg of concentrated supernatant protein was loaded.