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Switching from Epo to SCF induced a marked increase in Kit level (Figure 3A).
In this regard, SCF and Epo cooperation would be restricted to early stages of erythropoiesis when Kit level is high.
As illustrated in Figure 5B, expression of MT-LynY397F detected by immunoblotting with the MT antibody was associated with an increase in Kit level in the Epo-cultured cells.
The high Kit level in SCF-cultured cells was reminiscent of a mechanism involving SCF as a modulator of its own receptor expression as described for IL-3, CSF-1 and GM-CSF [25].
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Reciprocally, Kit levels were decreased when cells were switched from SCF to Epo for 48 hrs (Figure 3B).
Clearly, Kit levels were high in cells exhibiting a robust proliferative response to SCF (cell lines 633 and 663).
Thus, we investigated whether Kit levels could be reversibly modulated in response to a change in cytokines.
Modifications in Kit levels were detected both in whole cell extracts and at the cell surface indicating that they in fine may lead to a modified response to SCF.
However, we observed that Kit levels were modulated by the cytokine used to expand the cells (Epo versus SCF) and that this modulation was rapidly reversible by a change in cytokine.
The rapid up or down modulation of Kit levels in response to cytokine changes, the absence of cell death (data not shown) during cytokine changes and the reversibility of the phenomenon indicated that they did not result from a selection process.
When miRNA treatment becomes available, targeted overexpression of members of the miR-221/222 and miR-17-92 miR-17-92 miR-17-92ght significlustersuppress ETV1 and Kin levels.
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