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Blood, Vol. 110, Issue 6, 1840-1847, September 15, 2007
Direct interaction between Kit and the interleukin-7 receptor
Blood Jahn et al.
110: 1840
Supplemental materials for Jahn et al, Vol. 110, Issue 6, 1840-1847
Files in this Data Supplement:
- Figure S1. Characterization of Jurkat-K7 cells and functionality of Kit and IL-7R receptors in Jurkat-K7 cells (JPG, 38.3 KB)
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(A) Flow cytometry analysis of Jurkat WT and Jurkat-K7 cells. Jurkat WT (interrupted lines) and lentivirally transduced Jurkat-K7 cells (solid lines) were analyzed for Kit and IL-7R surface expression by staining the cells with antibodies against Kit (allophycocyanin-conjugated APC, upper panel) and IL-7R (phycoerythrin-conjugated PE, lower panel). (B, C) Functionality of Kit and IL-7R pathways in Jurkat-K7 cells. Jurkat WT and Jurkat-K7 cells were stimulated with KL or IL-7. Cell lysates were analyzed for Kit-induced total tyrosine phosphorylation (using an antiphosphotyrosine (PY) antibody) and Kit protein (B, upper and lower panels, respectively). Immunoprecipitated Stat5 protein was analyzed for IL-7 induced tyrosine phosphorylation of Stat5 (Tyr 694, upper panel) and Stat5 protein (C, lower panel).
- Figure S2. In 293T cells, Kit-induced activation of Stat5 is independent of Jak3 (JPG, 33.4 KB)
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(A) IL-7--induced activation of Stat5 is dependent on Jak3. 293T cells expressing IL-7R in the presence (lanes 1,2) or absence of Jak3 (lanes 3,4) were stimulated with IL-7. Immunoprecipitated Stat5 protein was analyzed for tyrosine phosphorylation (upper panel) and Stat5 protein (lower panel). (B) KL-induced activation of Stat5b is independent of Jak3 in 293T cells. 293T cells co-expressing Kit and IL-7R (lanes 1-4) in the presence (lanes 1,2) or absence of Jak3 (lanes 3,4) or Kit alone (lanes 5,6) were stimulated with KL for five minutes and immunoprecipitated Stat5 protein was analyzed for tyrosine phosphorylation (upper panel) and Stat5 protein (lower panel).
- Figure S3. Kit-mediated activation of Stat5 is dependent on Tyr567 and Tyr569 of Kit (JPG, 23.3 KB)
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293T cells expressing KitWT or mutant Kit constructs as indicated were stimulated with KL and lysates were analyzed for Stat5 phosphorylation on Tyr694 (upper panel), total Stat5 (middle panel) and Kit protein (lower panel).
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