Fig. 6.

Ethanol does not significantly alter dynein or dynactin protein expression levels or membrane attachment but enhances dynein microtubule binding. A Whole cell lysates from control (con) or treated (EtOH) cells were immunoblotted for dynein intermediate chain (dyn) or p150. Relative levels were determined with densitometry and normalized to total α-tubulin. Values indicate the percent expression in ethanol-treated cells relative to control and are the mean ± SEM from at least four independent experiments. B Post-nuclear supernatants (PNS), nuclear pellets (nuc) and cytosolic (S3), heavy (P2), and light membrane (P3) fractions from control or ethanol-treated cells (see “Materials and methods” section) were immunoblotted for dynein intermediate chain (dyn), p150, or atubulin (tub). Representative immunoblots are shown. C Polymerized microtubules were pelleted from cytosolic extracts (see “Materials and methods” section) from control or ethanol (EtOH) treated cells. The supernatant (S) and microtubule pellet (P) were immunoblotted for dynein intermediate chain (dyn), p150, or α-tubulin (tub). Representative immunoblots are shown