Description

Movie raw file of RPE-1 cells expressing GFP-alpha-tubulin and depleted for pVHL (Von Hippel-Lindau) by ectopic expression of shRNAmir against VHL. Video used to manually track MTs and determine growth speed and shrinkage time. Cells on round coverslips were transferred to a homemade holding device, and 600 µl of the appropriate medium was added. The video was acquired with a microscope (IX70 Delta Vision Spectris; Olympus), temperature-controlled at 37C, using a 100× NA 1.4 differential interference contrast (DIC) oil Plan-Apochromat objective, ex 470 em 520, and a camera (CoolSNAP HQ; Roper Industries) with an exposure time of 200 ms and a frame rate of 2Hz. Acquisition software used was SoftWoRx version 3.3.4 (Applied Precision). This video corresponds to J Cell Biol. 2010. 190: 991-1003--Supplementary Figure 1.