Figure 4. Colocalization effect of overexpressed PML and LC3 during cell cycle.

PC3 cells were transiently transfected with DsRed-PML IV and GFP-LC3 expressing plasmids and followed by treatment with nocodazole at 200 ng/ml for 18 hours. The treated cells were harvested by mitotic shake off, seeded onto coverslips and released from nocodazole block for 1 and 2 hours. Then the cells were fixed after staining with hoechest 33342 (blue) for observation with confocal microscopy. Representative images of dividing and G1 phase cells were observed (scale bar  = 10 µM). Right graph presented line scan analysis for colocalization of DsRed-PML and GFP-LC3 based on fluorescence intensity profiles across PML NBs as indicated on left merged images. Experiments were repeated for three times and similar results were obtained.