Christopher S. Wood, Karl R. Schmitz, Nicholas J. Bessman, Thanuja Gangi Setty, Kathryn M. Ferguson, Christopher G. Burd (2011) CIL:13450, Saccharomyces cerevisiae S288c. CIL. Dataset

Kre2-GFP is efficiently maintained in the Golgi apparatus in BY4742 vps74∆ expressing wild-type, untagged, Vps74. Control image for images showing that localization of Kre2-GFP is lost when mutant forms of Vps74 (mutations in the sulfate-binding pocket) are expressed (CIL#s 13452, 13454). 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 Fig4C top panel in J Cell Biol. 187: 967-975. 2009. Images in Fig 4C include CIL#s 13450, 13452, 13454.

Kre2-GFP 在 BY4742 vps74∆ 表达野生型、未标记的 Vps74 的 Golgi 细胞器中得以高效维持。控制图像显示，当表达 Vps74 的突变体（硫酸结合口袋中的突变）时，Kre2-GFP 的定位将丢失（CIL#s 13452, 13454）。本研究证实，Vps74 和 GOLPH3 的硫酸结合口袋介导 PtdIns4P 结合，并对于其功能至关重要。在液体培养基中培养的细胞被固定于生长培养基中，并在 DeltaVision 工作站（Applied Precision）上基于倒置显微镜（IX-70; Olympus）使用 100× NA 1.4 油浸镜头，以 0.4-µm 的 z 轴间距收集 3D 图像栈。图像在 23°C 下使用 12 位 CCD 相机（CoolSnap HQ; Photometrics）捕获，并利用迭代约束算法（Agard, 1984）以及测量的点扩散函数进行去卷积。J Cell Biol. 187: 967-975. 2009 年发表的文章图 4C 顶部面板中展示了一张 z 栈中心的图像。图 4C 中的图像包括 CIL#s 13450, 13452, 13454。