Figure 6

mRNA localization in subcellular foci. HeLa cells were observed using confocal microscopy 3 h, 18 h and 48 h post-transfection with 500 ng of red fluorescent-labeled V5PB mRNA polyplexes. Experiments have been done three times. GE-1/Hedls (P bodies marker) (A) and the eukaryotic initiation factor-3 eIF3η (Stress Granules marker, SGs) (B) were immunodetected using specific green fluorescent secondary antibodies. Insets show enlargements of the boxed areas with separated colors. Arrows indicate representative foci. The primary antibody directed against the nuclear p70 S6 kinase protein reveals P bodies foci due to cross-reactivity with GE-1/Hedls, and exhibits nuclear staining due to p70 S6 kinase protein detection. Nuclei were stained in green (A) or blue by DRAQ5® (B). BF: Bright Field. The images shown are representative Z-section from three separated experiments. No re5d fluorescent mRNA was observed in nuclei.