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:13907, 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 treated with 100 µM resveratrol for 4 h. 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 middle panel, in Morselli et al. J Cell Biol 192: 615-629

人类结肠癌细胞系HCT 116经过SIRT1特异性的siRNA转染后，再以编码GFP-LC3的质粒进行重转染，于全培养基中培养24小时，随后以100 µM的.resveratrol处理4小时。为进行荧光显微镜检测，将培养于载玻片上的细胞用4% wt/vol的甲醛在室温下固定15分钟，随后在磷酸盐缓冲溶液（PBS）中洗涤三次，并使用 mounting medium（Vectashield）进行封片。采用配置有阿贝式63× 1.3 NA浸没物镜的共聚焦荧光显微镜（TCS SP2；Leica）捕获共聚焦荧光图像。图像通过配置有DFC 350 FX 1.8.0相机（Leica）的LAS AF软件（Leica）获取，并使用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对比度和亮度的线性调整，操作均采用Photoshop（所有图片调整参数保持一致）；未使用显式的γ校正。图像：图1A，底部中间面板，见Morselli等人发表的《细胞生物学杂志》第192卷第615-629页。