Bartonella effector protein C mediates actin stress fiber formation via recruitment of GEF-H1 to the plasma membrane
Fig 2
The BepCBhe FIC domain but not a conserved FIC motif or flap region is required for actin stress fiber formation in B. henselae-infected HeLa cells.
(A) HeLa cells were infected with isogenic Bhe ΔbepA-G strains expressing 3xFLAG-tagged BepCBhe wild-type or mutant versions or carrying the empty plasmid at a multiplicity of infection (MOI) of 400. After 48 h of infection, cells were fixed and immunocytochemically stained, followed by fluorescence microscopy analysis. F-actin is represented in green, DNA in blue, and bacteria in red (scale bar = 50 μm). (B) HeLa cells were infected with the same strains as in (A) at MOI 50, 100, 200, 400, and 800, fixed after 24 h and 48 h and stained for F-actin. The graphs show the relative mean fluorescence intensity of the F-actin signal at 24 h (left panel) and 48 h (right panel) infection for the indicated MOIs normalized to the uninfected control. Shown are representative results from three independent experiments. BepCBhe**** = BepCBhe H146A, K150A, R154A, R157A; BepCBhe (Flap BepABhe) = BepCBhe A90E, R92K, P93R, K94T, H96W, R97K, V98N, P99A.