FIG. 2.

YE3614 is secreted into culture supernatants. (A) pSW43 (JB580v yspM ORF and promoter) was transformed into the wild-type (WT; JB580v), ΔysaC (YVM1178), and ΔyscC (YVM1374) strains. Bacteria were grown under Ysa T3SS-inducing conditions. Samples were harvested after 6 h, and samples were prepared for Western analysis with anti-CyaA antibody as described in Materials and Methods. (B) pSW41 (JB580v yspM ORF and yopE promoter) was transformed into the strains listed above. Bacteria were then grown under Ysc T3SS-inducing conditions (37°C and absence of Ca²⁺), and samples were prepared as described in panel A. (C) pSW45 (Y295 yspM ORF and promoter) was transformed into the strains listed above and grown under Ysa T3SS-inducing conditions, and supernatants and cells were analyzed as described in panel A.