Fig 4. Rab21CA over-expression is associated with increased invasion and matrix degradation.

A. Trophozoites stably expressing Rab21WT, Rab21KD, Rab21CA and Rab21DN and vector control (pEhExHA for Rab21WT and mutants; psAP2Gunma for Rab21KD strain) were harvested, washed and resuspended in serum free BI medium and 7.5 x10⁵ cells were plated in upper chamber of a transwell insert coated with 5mg/ml of matrigel. The cells were attracted in the lower chamber using 15% adult bovine serum as a chemoattractant. Trophozoites were allowed to invade for 48–50hrs.After 48hrs, the migrated cells in the lower chamber were detached and counted. Graph shows mean ± SEM of three independent experiments. B. Trophozoites stably expressing Rab21CA and Rab21DN were plated on glass coverslips coated with Hilyte488 Fibronectin and incubated at 35°C for 45–48 hrs under dark. After 48hrs, the cells were fixed and stained with Alexa568 Phalloidin for actin and confocal images were acquired using Zeiss LSM780. Scale bars, 10μm.