Fig. 5

A defect in c-Abl signaling contributes to the impairment of Hem-1^−/− HSCs. a As expected, depletion of Hem-1 protein results in degradation of other WAVE2 components such as Abi-1, Abi-2, WAVE2, and Sra-1. Expression of c-Abl protein was also decreased in Hem-1^−/− E14.5 CD45⁺ Lin⁻ FL cells as measured by western blot. β-Actin was used as a loading control. Optical density (OD) averages of c-Abl signal normalized to actin (±S.D.) of three distinct western analyses (n = 3, ***p < 0.001, Student’s t test). b Flow cytometric analysis of the phosphorylation of downstream effectors of the c-Abl signaling pathway in E14.5 CD45⁺ Lin⁻ FL cells found no differences compared to littermate Hem-1^+/+ cells (n = 9). c Quantitative RT-PCR expression analysis of regulators of apoptosis between Hem-1^+/+ and littermate Hem-1^−/− E14.5 CD45⁺ Lin⁻ FL (n = 9, **p < 0.01, Student’s t test). d The phosphorylation of c-Abl downstream signaling pathway effectors in Hem-1^−/− PD3 Lin⁻ BM cells was reduced compared to littermate Hem-1^+/+ equivalent cells (n = 9, *p < 0.05, **p < 0.01, ***p < 0.001, Student’s t test). e, Quantitative RT-PCR expression analysis of regulators of apoptosis found that pro-apoptotic effectors were increased and anti-apoptotic effectors were decreased in Hem-1^−/− PD3 Lin⁻ BM cells compared to littermate Hem-1^+/+ equivalent cells (n = 9, **p < 0.01, ***p < 0.001, ****p < 0.0001, Student’s t test)