American Society of Hematology

Figure 3.

$Figure 3. IL-6 induced translocation of CD45 to lipid rafts. (A) Lysates of U266 CD45+ or ILKM2 cells were separated by sucrose density gradient centrifugation, and continuous fractions (fractions 1 to 12) were examined by immunoblotting. Raft fractions were identified by dot-blotting with horseradish peroxidase-conjugated CTB. (B) For the non-cross-linked condition, cells were fixed and then stained with antibody (Ab) against CD45RO, CD45RB, IL-6Rα, or CD71, and subsequently with rhodamine-conjugated secondary antibody. Lipid rafts were stained with FITC-labeled CTB. Representative images were obtained by LCM. For the cross-linking condition, lipid rafts were pretreated with CTB-FITC followed by anti-CTB monoclonal antibody. Then, treated cells were fixed and stained similarly to panel A except for CTB staining.$

IL-6 induced translocation of CD45 to lipid rafts. (A) Lysates of U266 CD45⁺ or ILKM2 cells were separated by sucrose density gradient centrifugation, and continuous fractions (fractions 1 to 12) were examined by immunoblotting. Raft fractions were identified by dot-blotting with horseradish peroxidase-conjugated CTB. (B) For the non-cross-linked condition, cells were fixed and then stained with antibody (Ab) against CD45RO, CD45RB, IL-6Rα, or CD71, and subsequently with rhodamine-conjugated secondary antibody. Lipid rafts were stained with FITC-labeled CTB. Representative images were obtained by LCM. For the cross-linking condition, lipid rafts were pretreated with CTB-FITC followed by anti-CTB monoclonal antibody. Then, treated cells were fixed and stained similarly to panel A except for CTB staining.

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