Detection and monitoring of the secretion of the N-terminal fragment of enolase conjugated to EGFP and FLAG. (A) Detection of secreted enolase fragments. Cont., pUL-ATG-EGFP; ALL, pULGI2-ENO2; (1–17), pUL-eno(1–17); (1–28), pUL-eno(1–28); (1–30), pUL-eno(1–30); (1–50), pUL-eno(1–50); (1–110), pUL-eno(1–110); (1–169), pUL-eno(1–169). (B) SEC23-independent secretion of the enolase fragment. Secretion levels of recombinant proteins in sec23-1 strains under 25°C or 37°C are shown. (1–28), pUL-eno(1–28); ALL, pULGI2-ENO2; 37, cultivated at 37°C; 25, cultivated at 25°C. (C) Fluorescence microscopy of cells transformed with pUL-eno(1–28), pULGI2-ENO2, and pULGI2-PGI1. Bar, 10 μm. Similar results were obtained from 3 independent experiments.