RCC-4 cell stained with anti-alpha-tubulin (green) and anti-GTP-tubulin (red) antibodies. GTP-tubulin localizes to the microtubule (MT) tip and remnants of GTP-tubulin are found along the entire MT lattice. RCC-4 cells lack Von Hippel-Lindau (VHL) and when pVHL30 is expressed in these cells, a higher frequency of GTP caps and GTP remnants is observed. RCC-4 cells were permeabilized in PEM buffer with glycerol and 0.1% Triton-X100, then incubated with primary antibody hMB11 followed by incubation with secondary antibody. Cells were then fixed in cold methanol for 5 min and stained with the polyclonal anti-tubulin antibody followed by secondary antibody. Stacks of images were acquired with an IX70 Delta Vision Spectris microscope using a 60X NA1.4 DIC oil Plan-Apochromat objective. This stack corresponds to J Cell Biol. 2010. 190: 991-1003--control Fig 5F (left 2 panels) and the experimental with ectopic pVHL30 (right 2 panels in Fig 5F) is CIL# 26572.
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