A streptococcal Fic domain-containing protein disrupts blood-brain barrier integrity by activating moesin in endothelial cells
Fig 5
BifA leads to moesin T558 phosphorylation.
(A-D) Western blot detection of moesin phosphorylation upon BifA treatment. Human BMEC cells were incubated with different BifA variants for the indicated times and lysates were probed with appropriate antibody. GAPDH is a loading control. The graphs on the right panel show the quantitation of the moesin and phospho-moesin bands by ImageJ, normalized with GAPDH. (E) Effect of PKC inhibition upon BifA induction of moesin phosphorylation. Cells were pre-treated with PKC inhibitor NSC305787 for 30 min before BifA treatment, then analyzed by immunoblot with phospho-moesin antibody. (F) BifA treatment leads to formation of GTP-RhoA. Western blots were performed on lysates of cells treated with BifA alone, or BifA in the presence of NSC305787. Total RhoA and rhotekin protein precipitated GTP-RhoA was detected by anti-RhoA antibody. GAPDH was the loading control.