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Fig. 3 | Molecular Medicine

Fig. 3

From: The βc Component of the Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF)/Interleukin 3 (IL-3)/IL-5 Receptor Interacts with a Hybrid GM-CSF/Erythropoietin Receptor to Influence Proliferation and β-Globin mRNA Expression

Fig. 3

Tyrosine phosphorylation of receptor subunits

(A) Double immunoprecipitation of Ba/F3-GMER + βc cells before (0) and after (1) hGM-CSF stimulation. Anti-phosphotyrosine immunoprecipitates were subjected to a second immunoprecipitation with rabbit polyclonal antibodies to the C terminus of βc. Background phosphorylation is increased after hGM-CSF stimulation, similar to control Ba/F3-GMRα + βc cells. (B) Immunoprecipitation/Western analysis of Ba/F3-GMER (1), Ba/F3-GMER + βc (2), and Ba/F3-GMER + βtr (3) before (0) and after (1) stimulation with 0.1 or 20 nM hGM-CSF. Cellular proteins were immunoprecipitated with 4G10 antibodies to phosphotyrosine, subjected to SDS polyacrylamide gel electrophoresis, and transferred to a nylon membrane. Western analysis was then carried out using antibodies to the C terminus of the mEpoR. A dose-related induction of phosphorylation was seen in each case, similar to control Ba/F3-EpoR cells stimulated with Epo, shown on the right. These data are from one of three representative experiments.

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Molecular Medicine

ISSN: 1528-3658