Immunofluorescent analysis of pro-and anti-apoptotic proteins. Immunofluorescent analysis of the expression of Bax (green), Ku70 (green), and Sirt1 (red) proteins in E1A.NR3 retinal cells in the presence and absence of 40 μM resveratrol pretreatment (16 hrs) followed by induction of apoptosis using 0.8 mg/ml of Rec-1 or Enol-1 for 2 hrs. The cells were incubated with anti-Bax, anti-Ku70 and anti-Sirt1 antibodies followed by incubation in appropriate fluorescent secondary antibodies. Nuclei were stained with DAPI (blue) for 10 min. The images were photographed under a Leica DM5000B fluorescence microscope. Each treatment was replicated three times and the experiment was repeated twice. The arrows indicate the expression of corresponding proteins in the cytoplasm of nucleus.