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. 2014 Feb 20;111(10):E953–E961. doi: 10.1073/pnas.1322889111

Fig. 3.

Fig. 3.

Membrane binding of TsaP depends on PilQ. (A) Immunoblot analysis of equal amounts of total cell extracts of the WT, ΔpilQ, ΔtsaP, and ΔtsaP/tsaP+ strains grown in the presence of 1 mM IPTG using α-TsaP and α-PilE antibodies. (B) Upper part of Coomassie-stained SDS/PAGE of nonphenol-treated OM fractions isolated from the indicated N. gonorrhoeae strains. (C, Left) Coomassie-stained SDS/PAGE of phenol-treated membrane fractions from the indicated strains. (C, Right) Immunoblot analysis of the same samples using α-TsaP, α-PilQ, and α-PilE antibodies. (D) Total membranes (TM) derived from N. gonorrhoeae WT (Left) and the ΔpilQ mutant (Right) were treated twice for 30 min with 7.5 M urea. After centrifugation, the supernatants (W1 and W2) and the resuspended membrane pellets (P) were analyzed by immunoblot analysis using α-TsaP antibodies.