Figure 1.

Plcγ-1 is a direct downstream target of EpoR–Jak2 signaling. (a) Western blot analysis of Ba/F3 and 32D cells stably transfected with EpoR and Jak2-WT and of the erythroid progenitor cell line I/11. Analysis was performed using phospho (p) and total antibodies. These data are representative results of three independent experiments. (b) Kinetic analysis of the erythroid progenitor cell line I/11. These blots are representative results of three independent experiments. (c) Western blot analysis of cells treated with Actinomycin D (2 h; 0.5, 1 μg/ml). These data are representative results of three independent experiments. (d) Co-immunoprecipitation of Plcγ-1 and EpoR in erythroid progenitor cells (I/11). Immunoprecipitation was performed with antibodies specific for Plcγ-1. Western blotting of the precipitate was performed with anti-EpoR antibodies. These data are representative results of three independent experiments. (e and f) Western blot analysis of Ba/F3 and 32D cells stably transfected with EpoR and Jak2-WT and of the erythroid progenitor cell line I/11. Cells were treated with LY292004 (1 h; 100 μM) and U0126 (1 h; 5 μM), respectively. These blots are representative results of three independent experiments