Figure 5. RpRalF modulates actin dynamics.

A) Hela cells transfected with plasmids encoding YFP, YFP-RpRalF_1–458 or YFP-RpRalF_1–458E100A were fixed 24 hours after transfection, then stained with Texas Red Phalloidin. Bar = 10 µm. B) Actin stress fibers disruption quantification. Cells were transfected with indicated YFP-tagged RpRalF or LpRalF constructs. 24 h after transfection, cells were fixed and actin was stained with phalloidin. The proportion of cells containing stress fibers was quantified for each construct. The standard deviation is derived from 3 independent experiments (* P<0.01, compared to YFP alone). C) and D) The RpRalF capping domain and a proline-rich region direct effector functions. C) Localization of ectopically expressed RpRalF_189–458, RpRalF_189–359 and RpRalF_359–458. Hela cells were transfected with YFP-tagged RpRalF_189–458, RpRalF_189–359 or RpRalF_359–458. 24 h after transfection, cells were fixed and stained with Texas Red phalloidin. Bar = 10 µm. D) RpRalF_189–359 and RpRalF_359–458 domains are both required for stress fibers disruption. Cells were transfected with indicated YFP-tagged RpRalF constructs. 24 h after transfection, cells were fixed and actin was stained with phalloidin. The proportion of cells containing stress fibers was quantified for each construct. The standard deviation is derived from 3 independent experiments (* * P<0.001).