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IL-3–induced enhancement of retinoic acid receptor activity is mediated through Stat5, which physically associates with retinoic acid receptors in an IL-3– dependent manner by Jutong Si, and Steven J. Collins Blood Volume 100(13):4401-4409 December 15, 2002 ©2002 by American Society of Hematology SCF activates the MAP kinase and PI3 kinase signaling pathways in cultured EML cells.EML cells were washed and cultured for 4 hours in SCF-deficient media. Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology IL-3 activates multiple signal transduction pathways in the SCF-dependent EML cells.IL-3 (5 ng/mL) was added to EML cells actively proliferating in SCF-containing media. Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology Effect of chemical inhibitors on the IL-3–induced activation of different signal transduction pathways in the SCF-dependent EML cells.EML cells cultured in SCF-containing media were incubated for 60 minutes in (A) the MEK1/2 inhibitors PD98059 and U0126 (la... Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology The Jak2 inhibitor (AG490) inhibits the IL-3–induced enhancement of RAR transcriptional activity in EML cells.The SCF-dependent EML cells were incubated overnight (15 hours) in the presence or absence of IL-3 (5 ng/mL) as indicated. Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology Activated Stat5 directly mediates the IL-3 induction of enhanced RAR transcriptional activity in EML cells.(A) Uninduced EML cells were electroporated with the βRARE-tk-Luc reporter (25 μg) together with expression vectors (40 μg of each) for wild-type Stat... Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology Constitutively active Stat5 enhances CFU-GM generation in SCF-dependent EML cells.EML cells were electroporated with different expression vectors including an empty vector, the wild-type Stat 5 (Stat5aWT), or the constitutively active Stat5 (Stat5aHS; 40 μg... Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology A Stat5-binding site is present in the RARE within the RARβ promoter.(A) Sequence of the RARE within the RARβ promoter (−55 to −33).21 The two 6-bp direct repeats making up the DR5 element are in bold. Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology Stat5 associates with RXR-RAR in an IL-3–dependent manner.(A) Cellular lysates (2 mg) of BAF3 cells that had been stably transduced with a FLAG-tagged RXR cDNA (BaF3-RXR cells) were immunoprecipitated with either Stat5 antibody (lane 3) or control IgG (lane... Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology Stat5 is likely not an immediate early activator of the βRARE.(A) SCF-dependent EML cells were incubated with IL-3 for the indicated period of time, and the cells were then electroporated with the βRARE-tk-Luc reporter (25 μg). Jutong Si, and Steven J. Collins Blood 2002;100:4401-4409 ©2002 by American Society of Hematology
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