Fig 4.
Infections by rOC/ATCC and rOC/US183-241 promote BAX, CytC, and AIF relocalization in human neurons. Differentiated LA-N-5 cells were infected with rOC/ATCC or rOC/US183-241. (A) Immunofluorescent detection of activated BAX. Cells were incubated with the MitoTracker Red CMXROS (red), fixed, and incubated with an anti-activated BAX antibody (green). Colocalization is represented by merged BAX and MitoTracker Red CMXROS signals (yellow) as indicated by white arrows. (B) Western immunoblotting of mitochondrial BAX and CytC. Mitochondrial protein fractions were subjected to Western immunoblotting analysis using antibodies directed against BAX or CytC. VDAC served as a loading control. (C) Immunofluorescent detection of AIF. Cells were incubated with the MitoTracker Red CMXROS (red), fixed, and incubated with an anti-AIF antibody (green) and DRAQ5 (blue). Nuclear colocalization is represented by merged AIF and DRAQ5 signals (turquoise) as indicated by white arrows, and residual mitochondrial colocalization is represented by merged AIF and MitoTracker (yellow) as indicated by white arrowheads. (D) Western immunoblotting of nuclear AIF. Nuclear protein fractions were subjected to Western immunoblotting analysis using antibodies directed against AIF. p84 served as a loading control.