FIG. 2.

AvrPto1-FLAG is secreted by hrpG, hrpJ, hrpV, and hrpT mutants in P. syringae pv. syringae 61. The indicated wild-type, hrp mutant, or ΔhrcC strains were grown in Hrp-inducing minimal medium to an optical density at 600 nm of 0.3. Cultures were centrifuged to separate cell pellet (C) and supernatant (S) fractions. Fractions were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane. AvrPto1-FLAG and NptII were detected by immunoblotting with either anti-FLAG or anti-NPTII antibodies, followed by secondary antibodies conjugated to alkaline phosphatase. NptII is a cytoplasmic protein used as a control for cell lysis.