Human colon carcinoma HCT 116 cells were transfected with a GFP-LC3-encoding plasmid, cultured in complete medium for 24 h and enucleated to obtain cytoplasts. The cytoplasts were treated with 100 µM Resveratrol for 4 h. Cytoplasts were still able to accumulate GFP-LC3 puncta in response to spermidine or resveratrol treatment, indicating that nuclei (and by extension transcription) are not required for short-term autophagy stimulation by these two agents. Nuclei were counterstained by Hoechst. Confocal fluorescent images were captured using a confocal fluorescence microscope (TCS SP2; Leica) fitted with an Apochromat 63× 1.3 NA immersion objective. Images were acquired with a camera (DFC 350 FX 1.8.0; Leica) using LAS AF software (Leica) and processed with Photoshop (CS2; Adobe) software. Specifically, picture processing involved cropping of representative areas and linear adjustments of contrast and brightness and was performed using Photoshop (with equal adjustment parameters for all pictures); no explicit γ correction was used. Image: Figure 8A, middle panel, in Morselli et al. J Cell Biol 192: 615-629