Fig 7. trans-Sialidase is shed in microvesicles.

Western blots of microvesicles purified by two independent methods, Exoquick kit (A) and ultracentrifugation (B), show TS presence only in the microvesicles fraction. Mucins were found predominantly in microvesicles although also to a minor extent in the supernatant. TolT and CLCP proteins contained in DRMs, were not found in the conditioned media (A). TS was hydrolyzed by the addition of recombinant PI-PLC (rPI-PLC) from microvesicles (C) and from trypomastigotes (D) confirming the absence of endogenous PI-PLC. TS was also found in microvesicles from the supernatant of infected cell cultures evidencing no involvement of PI-PLC activity from Vero cells or amastigotes (E). F) and G) are TEM micrographs of microvesicles purified by Exoquick and ultracentrifugation respectively. Try; trypomastigotes, CM; conditioned medium, μV; microvesicles; SN; supernatant.