SFK and c-KIT knockdown using siRNA inhibits survival and growth of primitive and committed AML progenitors. (A) LYN, HCK, FGR, and c-KIT gene expression fold change in AML CD34⁺ cells (n = 5; ***P < .001) (left). (right) Detection of SFKs phosphorylation by flow cytometry 72 hours postnucleofection (n = 5; **P < .01). Results shown represent the mean ± SEM of 5 AML samples. (B) AML CD34⁺ apoptosis (n = 5) 72 hours postnucleofection. Results are presented as mean ± SEM of Annexin V-positive cells for 5 AML samples. ns, non significant; **P < .01. (C) CFC assay 72 hours postnucleofection (n = 4). Results are presented as percentage of control and are mean ± SEM for 4 AML samples. *P < .05; **P < .001; ***P < .001. (D) AML CD34⁺ apoptosis (n = 5; ***P < .001) 72 hours postnucleofection with indicated siRNA. Results represent mean ± SEM of Annexin V-positive cells. ns, non significant; ***P < .001. (E) CFC assay 72 hours postnucleofection (n = 5). Results shown are percentage of control and represent mean ± SEM. **P < .01; ***P < .001. (F) SFK phosphorylation 2 hours after dasatinib treatment (200 nM) for AML CD34⁺CD38⁻ (left; *P < .05) and AML CD34⁺CD38⁺ cells (right; *P < .05). Results are representative of 6 AML samples. (G-H) Western blot analysis of c-KIT and SFK phosphorylation in CD34⁺ cells from 3 AML patients cultured for 2 hours (G) or 48 hours (H) without or with 200 nM dasatinib. Results shown are representative of 6 AML samples.