Description

Immunofluorescence images used to investigate stabilization of microtubules (MTs) by Von Hippel-Lindau tumor suppressor protein (pVHL). VHL-deficient RCC-4 cells expressing wild-type VHL30 protein have a higher alpha-tubulin (MTs; green) signal relative to GSK-3beta (red; an internal fluorescence control), compared with RCC-4 control cells, suggesting that VHL stabilizes MTs. RCC-4 cells were infected with a retroviral vector to express pVHL30, fixed in 3% paraformaledehyde and permeabilized in 0.2% Triton X-100. Primary antibodies used were: monoclonal anti-GSK3beta and polyclonal anti-alpha tubulin. Secondary antibodies used were: anti-rabbit FITC and anti-mouse Texas-Red. Stacks of 5 z planes were acquired with a Leica SP2 system using the 63x 1.4NA, Oil, HCX Plan-Apo Objective and the distance between z planes is 1 micron. This stack corresponds to J Cell Biol. 2010. 190: 991-1003--Fig 1C (right panels) and the images of the control are in Fig 1C (left panels) and CIL# 31263.