First Characterization of the Neospora caninum Dense Granule Protein GRA9

Figure 8

The secretion of the HA-tagged NcGRA9 variants was analysed over time (5 min, 20 min, 2 h, 4 h, 8 h, and 24 h postinfection) by confocal microscopy. (a) shows that full-length NcGRA9-HA was present in the vacuolar space after 20 min (white arrow). After 4 h the protein was in the deeper vacuolar space (white arrow) and after 8 h full-length NcGRA9-HA was also present near the PVM (white arrow). (b) shows that 280 NcGRA9-HA secretion is delayed compared to full-length NcGRA9-HA and starts 4 h after invasion (white arrow). (c) shows that 201 NcGRA9-HA was retained at all analysed time points inside the tachyzoites in the paranuclear space (white arrows).Red: anti-HA cy3; blue: DAPI; merge: anti-HA, DAPI, DIC.

(a) N. caninum  +  NcGRA9-HA

(b) N. caninum  +  280 NcGRA9-HA

(c) N. caninum  +  201 NcGRA9-HA