Figure 7.
Association of newly synthesized AE1 with cytoskeletal ankyrin is inhibited by BFA. Erythroid cells from a 10-d-old chicken embryo were pulsed for 15 min with 35S-Translabel and chased for 1 h (lanes 1 and 2) or 4 h (lanes 3 and 4) in complete media. Alternatively, BFA was added to the media at the 45-min chase point and included in the media for the duration of a 4-h chase (lanes 5 and 6). At each time point, the cells were detergent lysed, and ankyrin immunoprecipitates were prepared from the detergent-soluble (S) and -insoluble (I) fractions. The ankyrin immunoprecipitates were dissociated in SDS sample buffer and reprecipitated with AE1-specific antibodies. The AE1 immunoprecipitates were analyzed on a 6% SDS polyacrylamide gel, and labeled species were detected by fluorography.