Eugenia Morselli, Guillermo Mariño, Martin v. Bennetzen, Tobias Eisenberg, Evgenia Megalou, Sabrina Schroeder, Sandra Cabrera, Paule Bénit, Pierre Rustin, Alfredo Criollo, Oliver Kepp, Lorenzo Galluzzi, Shensi Shen, Shoaib Ahmad Malik, Maria Chiara Maiuri, Yoshiyuki Horio, Carlos López-Otín, Jens S. Andersen, Nektarios Tavernarakis, Frank Madeo, Guido Kroemer (2011) CIL:13906, Homo sapiens, permanent cell line cell. CIL. Dataset

Human colon carcinoma HCT 116 cells were transfected with an siRNA specific for SIRT1 and then retransfected with a GFP-LC3-encoding plasmid, cultured in complete medium for 24 h and left untreated for 4 h as a control for resveratrol or spermidine treatment. For fluorescence microscopy determinations, cells cultured on coverslips were fixed in paraformaldehyde (4% wt/vol) for 15 min at RT, washed three times in PBS, and mounted with mounting medium (Vectashield). 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 1A, bottom left panel, in Morselli et al. J Cell Biol 192: 615-629

人结肠癌细胞HCT 116经SIRT1特异性siRNA转染后，再转染GFP-LC3编码质粒，于全培养基中培养24小时，并在4小时内未进行任何处理作为白藜芦醇或精胺处理的对照组。为进行荧光显微镜测定，将培养在盖玻片上的细胞用4% (wt/vol) 甲醛固定15分钟，室温下洗涤三次磷酸盐缓冲液（PBS），然后用封片剂（Vectashield）封片。采用共聚焦荧光显微镜（TCS SP2；莱卡）及配备阿贝63×1.3 NA浸没物镜进行共聚焦荧光图像的采集。图像通过莱卡DFC 350 FX 1.8.0相机（Leica）使用LAS AF软件（莱卡）获取，并经过Adobe Photoshop（CS2；Adobe）软件进行处理。具体而言，图像处理包括代表性区域的裁剪以及对比度和亮度的线性调整，使用Photoshop（所有图片具有相同的调整参数）进行；未使用显式γ校正。图像：Morselli等人《细胞生物学杂志》第192卷第615-629页图1A左下角板块。