Figure 3. Tubulin curls made with either... | Rockefeller University Press

Figure 3.

$Figure 3. Tubulin curls made with either GTP or GDP will pull down CENP-F better than Tx-MTs. (A) RPE1 cells were synchronized, collected, and extracted as described in Materials and methods, and then mixed with either vinblastine-induced tubulin curls or Tx-MTs and pelleted. A 4× pellet volume of cell extract and equal volumes of each pellet were fractionated on 7% polyacrylamide gels. 1, cell lysate; 2, 3, and 4, pellets of GTP curls, GDP curls, and Tx-MTs, respectively; 5, molecular mass markers. (B) The gels from A blotted to nitrocellulose paper and reacted with a mouse monoclonal antibody to CENP-F (Santa Cruz Biotechnology, Inc.). Numbers above blot lanes indicate the number of microliters loaded. (C) Ratios of curls/Tx-MTs staining, as measured by densitometry. The binding ratios for GTP curls and Tx-MTs shown here are from a single representative experiment out of three repeats.$

Tubulin curls made with either GTP or GDP will pull down CENP-F better than Tx-MTs. (A) RPE1 cells were synchronized, collected, and extracted as described in Materials and methods, and then mixed with either vinblastine-induced tubulin curls or Tx-MTs and pelleted. A 4× pellet volume of cell extract and equal volumes of each pellet were fractionated on 7% polyacrylamide gels. 1, cell lysate; 2, 3, and 4, pellets of GTP curls, GDP curls, and Tx-MTs, respectively; 5, molecular mass markers. (B) The gels from A blotted to nitrocellulose paper and reacted with a mouse monoclonal antibody to CENP-F (Santa Cruz Biotechnology, Inc.). Numbers above blot lanes indicate the number of microliters loaded. (C) Ratios of curls/Tx-MTs staining, as measured by densitometry. The binding ratios for GTP curls and Tx-MTs shown here are from a single representative experiment out of three repeats.