FIG. 3.

Expression of EGFP fusion proteins in HeLa cell after transient transfection. Wild type and exoS, exoS(E381A), and exoS(R146K/E381A) mutants were cloned into pEGFP-C1, with the resulting fusion constructs labeled as follows: S, wild-type ExoS; EA, ExoSE318A; and RKEA, ExoSR146KE381A double mutant. At 12 h posttransfection, the total cellular proteins were collected. The expression of EGFP or fusion proteins was determined by Western blotting with antibodies to ExoS or GFP. Arrows point to the full-length fusion protein band; the arrowhead indicates the EGFP band. Equal protein loading was controlled by Ponceau S staining of membranes and Western blotting for β-actin.