American Society of Hematology

Fig. 5.

$Fig. 5. P-gp association with ERM proteins. / (A) Upper panel: Western blotting for P-gp (arrow) in actin, ezrin, radixin, and moesin immunoprecipitates (IP) from the cytoskeletal/membrane fraction of CCRF-CEM (1) and CEM-VBL100 (2) cells. P-gp is clearly detectable in each ezrin, radixin, moesin, and actin immunoprecipitate only from CEM-VBL100 cells. Western blotting for P-gp in a CD4 immunoprecipitate from the cytoskeletal/membrane fraction of the same cell types (CD4) was included as a negative control for P-gp coimmunoprecipitation. Lower panel: Western blotting for actin, ezrin, radixin, moesin, and CD4 in actin, ezrin, radixin, moesin, and CD4 immunoprecipitates, respectively, from the cytoskeletal/membrane fraction of CCRF-CEM (1) and CEM-VBL100 (2) cells. To avoid overlapping with Ig heavy chains, actin and CD4 immunoprecipitates were separated by SDS-PAGE in nonreducing conditions (see “Materials and methods”), followed by blotting with the anti-actin or the anti CD4 mAb, respectively. The figure shows successful immunoprecipitation of each tested protein. (B) Western blotting for ezrin, radixin, and moesin in P-gp immunoprecipitates from the cytoskeletal/membrane fraction of CCRF-CEM (1) and CEM-VBL100 (2) cells. Each ERM protein is clearly detectable in P-gp immunoprecipitates only from CEM-VBL100 cell protein extracts.$

P-gp association with ERM proteins.

(A) Upper panel: Western blotting for P-gp (arrow) in actin, ezrin, radixin, and moesin immunoprecipitates (IP) from the cytoskeletal/membrane fraction of CCRF-CEM (1) and CEM-VBL100 (2) cells. P-gp is clearly detectable in each ezrin, radixin, moesin, and actin immunoprecipitate only from CEM-VBL100 cells. Western blotting for P-gp in a CD4 immunoprecipitate from the cytoskeletal/membrane fraction of the same cell types (CD4) was included as a negative control for P-gp coimmunoprecipitation. Lower panel: Western blotting for actin, ezrin, radixin, moesin, and CD4 in actin, ezrin, radixin, moesin, and CD4 immunoprecipitates, respectively, from the cytoskeletal/membrane fraction of CCRF-CEM (1) and CEM-VBL100 (2) cells. To avoid overlapping with Ig heavy chains, actin and CD4 immunoprecipitates were separated by SDS-PAGE in nonreducing conditions (see “Materials and methods”), followed by blotting with the anti-actin or the anti CD4 mAb, respectively. The figure shows successful immunoprecipitation of each tested protein. (B) Western blotting for ezrin, radixin, and moesin in P-gp immunoprecipitates from the cytoskeletal/membrane fraction of CCRF-CEM (1) and CEM-VBL100 (2) cells. Each ERM protein is clearly detectable in P-gp immunoprecipitates only from CEM-VBL100 cell protein extracts.

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