FIG 4.

Evaluation of C. trachomatis candidate secretion using a GSK FLAG secretion assay. Candidate secretion substrates were fused to a GSK FLAG tag and transformed into C. trachomatis. HeLa cells were infected at an MOI of 5 with each strain, and after 24 h, the candidate peptide was immunoprecipitated using anti-FLAG beads. Samples were resolved by SDS-PAGE, and immunoblots were probed with anti-GSK or anti-P-GSK to evaluate expression and secretion, respectively. CT149, CT131, CT627, and CT584 were determined to be secreted using this assay. The asterisk indicates the molecular weight of each GSK FLAG fusion. Predicted molecular weights are as follows: CT503-GSK-FLAG, 30 kDa; CT736-GSK-FLAG, 19 kDa; CT149-GSK-FLAG, 37 kDa; CT131-GSK-FLAG, 129 kDa; CT627-GSK-FLAG, 41 kDa; CT584-GSK-FLAG, 24 kDa. Data are representative of 3 independent experiments.