Dasatinib combined with chemotherapy enhances p53 transcriptional activity and modulates the Akt-HDM2 axis in AML CD34⁺ cells. (A) Quantitative PCR analysis of p53 and p53 target genes in primary AML CD34⁺ cells exposed for 16 hours to dasatinib alone (200 nM), DNR alone (50 nM), or dasatinib 200 nM in combination with DNR. β-2M was used as an internal control and results shown are expressed as mean ± SEM of 8 AML samples. One-way ANOVA with posttest: ns, nonsignificant; *P < .05; **P < .01; ***P < .001; ****P < .0001. (B) Western blot analysis for AML samples exposed for 48 hours to dasatinib alone (200 nM), DNR alone (50 nM), or dasatinib in combination with DNR. Indicated antibodies are listed and β-actin was used as a loading control. Results shown are representative of 7 AML samples analyzed. (C) Twenty-four hours postnucleofection, AML samples (n = 4) were treated without or with dasatinib (200 nM) + DNR (50 nM) and assayed for apoptosis after 2 more days. Results shown are presented as mean percentages ± SEM of Annexin V-positive cells in control versus p53 siRNA and in treated versus untreated cells. *P < .0201; **P < .0039; ***P < .0003. (D) Twenty-four hours postnucleofection with indicated siRNA, AML samples (n = 5) were treated without or with dasatinib (200 nM) + DNR (50 nM) for 16 hours and p53 gene expression was assessed using quantitative PCR analysis. β2M was used as an internal control; results shown are expressed as mean ± SEM of 5 AML samples. ns, nonsignificant; *P < .0278; **P < .0072. (E) Western blot analysis for AML samples exposed for 2 hours to dasatinib alone (200 nM), DNR alone (50 nM) or dasatinib in combination with DNR. Indicated antibodies are listed and β-actin was used as a loading control. Results shown are representative of 7 AML samples analyzed.