Immunofluorescence microscopy
Cells plated on cover slides were first incubated with 0.5 μM
Mitotracker Red (Invitrogen) at 37 °C for 30 min, and then fixed with 4
% formaldehyde. Following treatment with blocking buffer, the cells
were incubated with an eEF2 antibody at 4°C for overnight, followed by
incubation with the second antibodies, AX568 Fluor Red and Alexa Fluor
488 Green, at room temperature for 90 min. Finally, the sample slides
were mounted with ProLong Gold Antifade Reagent (Life Technologies) with
or without DAPI and incubated overnight at room temperature. The slides
were observed using a laser scanning confocal microscope (Nikon).