Fig. 2.

Time course of MT assembly/disassembly in Noco/Tri cells. 3T3 cells were pre-treated with 10 μM nocodazole prior to detergent extraction. Rat brain cytosol (25%) was then applied to the Noco/Tri cells in the presence of GTP to study MT assembly at various time points (a). Microtubule organizing center (MTOC)-like structures could be detected as early as 1 min after incubation with adult rat brain cytosol, whereas by 10 min, MT growth from MTOC could be observed. Assembled MTs were stable for up to 4 h studied. b To study disassembly, MTs were first assembled in Noco/Tri cells as described above for 20 min. The cells were then washed, incubated in PEM buffer, fixed at different time points, immunostained and analyzed by confocal microscopy. By 1 h, almost all MTs were disassembled. Anti-alpha tubulin (DM1A) was used for immunocytochemical staining. Scale bar represents 25 μm