Figure 9.

³²P-labeled moesin partitions with the Triton X-100 insoluble fraction. NIH3T3 cells were transfected with HA-moesin only or HA-moesin and RhoAV14 as indicated. Cells were then serum starved and metabolically labeled with [³²P]orthophosphate or [³⁵S]methionine/cysteine as described in Figure 8, and then subjected to 0.5% Triton X-100 detergent extraction. Soluble (S) and insoluble (I) fractions were immunoprecipitated with 12CA5 antibody. In contrast to total (³⁵S) moesin, ³²P-labeled moesin was found almost exclusively in the insoluble fraction. Also, coexpression of RhoAV14 increases the percentage of moesin in the insoluble fraction. The data shown are representative of three independent experiments.