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Figure 6


Fig. 6. Internalization and degradation of IL-2Rβ in BAF-B03 and its transfectants. (A) IL-2Rβ expression on the surface of the pro-B cell line clones F7, S25, BAFβd349-410-3 and BAFβd349-410-4 was examined by flow cytometry. Cells were incubated with an anti-IL-2Rβ monoclonal antibody (TU11), followed by a FITC-conjugated secondary antibody. (B) Aliquots (1.25%) of total lysates from the indicated BAF-B03 clones (2x106 cells) were immunoblotted with an anti-IL-2Rβ antibody (C-20) or anti-β-actin antibody. IB, immunoblotting. (C) Internalization of IL-2Rβ in the transfectants. The radioactivity of cell surface-bound acid-removable fractions (a) and intracellular acid-unremovable fractions (b) was counted. (D) Degradation of IL-2Rβ in the transfectants. The radioactivity of culture supernatants (a), cell precipitate fractions (b) and TCA-soluble fractions of culture supernatants (c) was counted. 125I-TU11 binding to parental BAF-B03 cells was 3.3% of that of F7 cells. The values represent the mean ± s.e.m. of triplicate determinations. Cells were incubated with (IL-2+) or without (IL-2–) IL-2.





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