Figure 3
From: Characterization of translocon proteins in the type III secretion system of Lawsonia intracellularis
LI1159 and LI1159 are the substrates of the Yersinia type 3 secretion system. T3SS-competent Y. enterocolitica MRS40∆YopHOPEM and T3SS-null Y. enterocolitica MRS40∆yscF expressing various HA-tagged proteins (LI1157, LI1158, LI1159, LI1158N40-YopE∆N15, LI1159N40-YopE∆N15, RplJ, YopE) were cultivated in BHI media under either T3SS-repressive (+ Ca2+) or -inductive (− Ca2+) conditions. The bacteria were induced to express the T3SS and transgenes by adding 0.2% L-arabinose and incubated at 37 °C for 6 h. Equal amounts of bacteria cultures were separated into cell-free culture supernatants (S) and whole-cell pellets (P), and samples corresponding to 0.10 OD620 per mL for S and 0.02 OD620 per mL for P were analyzed on 12% (w/v) polyacrylamide gels. Specific proteins were detected by immunoblot with anti-HA followed by visualization with alkaline phosphatase-conjugated secondary antibodies and development with ECL.