Subcellular localization of RGN-GFP in Cos-7 cells treated with Doxorubicin
Cos-7 cells transfected with RGN-GFP (green) were then treated with DOX (20 μM) for 4 h before being stained with mito-tracker (red) and Hoechst (blue) and then fixed with paraformaldehyde. The fixed cells were then viewed under a fluorescence microscope under (A) bright field illumination, to show morphological changes; (B) blue (nuclei); (C) green (RGN-GFP) channels and (D) red (mitochondria), to discriminate between the different locations. Panel (E) shows the superimposition of all three channels. Panel (F) shows the scatter plot of individual pixel intensities within the image for the green and red channels. Panel (G) shows the scatter plot of individual pixel intensities within the image for the green and blue channels. The images are presentative of three separate experiments, each analyzing several fields of view. The Pearson’s correlation coefficient between RGN and mitochondria was determined to be 0.8 and for RGN and nuclei it was 0.41, when analyzed for the three experiments.