Figure 6.
Differential partitioning of Gαi1 to the microtubule fraction in the taxol-resistant cells. The 2008, 2008/13/4 and 2008/17/4 cells were seeded at a density of 1 × 106 cells/plate and allowed to incubate in complete growth media for 36 hours. The cells were then treated with taxol (500 nM) for the indicated time. At the end of each time period, cells were processed for extraction of whole cell lysates (as described in Fig. 1), cytoskeletal fractions (as described in Fig. 5), cytoplasmic and nuclear fraction (utilizing the protocol supplied by manufacturer of the NE-PER extraction kit) and membrane fraction (utilizing the single step Mem-PER extraction kit). Protein (5 – 20 μg/lane) from each fraction were resolved on a 12% (w/v) SDS-PAG and then transferred to a PVDF membrane. Western blotting was performed utilizing the Gαi1 polyclonal antibody.