The c-Kit tyrosine kinase receptor and the SCL/Tal1 (Stem Cell Leukemia) transcription factor are crucial to hematopoiesis and are co-expressed by hematopoietic progenitors. In the present study, we directly address the question whether SCL and c-Kit establish a code for the fate of multipotent progenitors in the adult. We therefore proceeded to loss and gain of function experiments, via retroviral mediated gene transfer in primary hematopoietic cells. We show that SCL is functionally required to sustain the survival of c-Kit+ cells in response to Steel Factor (Kit ligand), but not to GM-CSF and IL-3. Interfering with SCL function using a DNA-binding defective SCL mutant (ΔbSCL) caused a 100-fold loss of multipotent progenitors, due to massive apoptotic death in the c-Kit+ fraction, indicating that the DNA binding function of SCL is essential in cell survival. An antisense SCL also caused apoptosis in c-Kit+ cells, which was rescued by co-delivering SCL in the sense orientation. Furthermore, SCL sets threshold for the response of multipotent progenitors to Steel factor without affecting their response to IL-3 or GM-CSF, demonstrating the specificity of SCL for c-Kit-dependent pathway. Moreover, SCL levels in purified hematopoietic progenitors (HSC, CMP, MEP, CLP) were 3.5 to 15-fold decreased in W41 W41 mice that have a hypomorphic c-Kit allele when compared to age-matched wild type controls. Conversely, ectopic SCL expression is sufficient to bypass c-Kit signalling to suppress apoptosis and increase the proliferative potential of multipotent progenitors, as anemia and other hematopoietic defects caused by a hypomorphic c-Kit allele were rescued by introducing an SCL transgene into the W41 W41 background. We conclude that SCL operates within the c-Kit pathway to suppress apoptosis and determine the clonal output of adult multipotent progenitors. To define the pathway through which SCL suppresses apoptosis in hematopoietic progenitors, we performed microarrays analysis on Δb-SCL expressing cells and identified novel survival genes that were confirmed to be direct SCL targets by chromatin immunoprecipitation. Finally, the expression of these target genes was decreased in purified progenitors from W41 W41 mice as assessed by quantitative PCR. Together, this study reveals a new and essential genetic pathway involving SCL downstream of c-Kit signalling for the survival of hematopoietic progenitors.