CIL:13449, Saccharomyces cerevisiae S288c. In Cell Image Library

Intracellular localization of GFP-Vps74 in BY4742 vps74∆. GFP-Vps74 is localized to Golgi compartments and the cytosol. Control image for images showing ectopic expression of GFP-Vps74 with mutations in the sulfate-binding pocket (CIL#s 13451, 13453). This study demonstrates that the sulfate-binding pocket of Vps74 and GOLPH3 mediates PtdIns4P-binding and is essential for function. Cells grown in liquid medium were mounted in growth medium and 3D image stacks were collected at 0.4-µm z increments on a DeltaVision workstation (Applied Precision) based on an inverted microscope (IX-70; Olympus) using a 100× NA 1.4 oil immersion lens. Images were captured at 23C with a 12-bit CCD camera (CoolSnap HQ; Photometrics) and deconvolved using the iterative-constrained algorithm (Agard, 1984) and the measured point spread function. One image from the approximate center of z stack is shown in Fig4A wild-type/vps74∆ panel in J Cell Biol. 187: 967-975. 2009. Images in Fig 4A include CIL#s 13449, 13451, 13453, 13455, 13456, 13457, 24816, 24817, 24818, 24819, 24820, 24821.

BY4742 vps74Δ菌株中GFP-Vps74的细胞内定位。GFP-Vps74定位于高尔基体区室与细胞质基质中。本图为显示硫酸结合口袋发生突变的GFP-Vps74异位表达图像的对照图像（CIL#s 13451、13453）。本研究证实，Vps74与GOLPH3的硫酸结合口袋可介导与磷脂酰肌醇4-磷酸（PtdIns4P）的结合，且该口袋对蛋白功能至关重要。将在液体培养基中培养的细胞置于生长培养基中封片，基于倒置显微镜（IX-70；奥林巴斯（Olympus））搭载的DeltaVision工作站（Applied Precision），使用100× NA 1.4油浸物镜，以0.4 μm的z轴步距采集三维图像堆栈。于23℃下使用12位CCD相机（CoolSnap HQ；Photometrics）采集图像，并采用迭代约束反卷积算法（Agard, 1984）与实测点扩散函数进行反卷积处理。选取z轴堆栈近似中心处的一幅图像，该图像见于《细胞生物学杂志》（*J Cell Biol*）2009年第187卷第967-975页的图4A野生型/vps74Δ组面板中。图4A中包含的图像对应CIL#s 13449、13451、13453、13455、13456、13457、24816、24817、24818、24819、24820、24821。