Figure 3

Comparison of canonical Ras effector and cancer signaling pathways in de novo NRD AML and relapsed NRI AMLs identifies potential mechanisms that drive relapse with NRI disease. (a) Flow cytometry histograms of Ras effector pathways (left) or BCL2 and BCL-xL protein levels (right) in splenocytes from leukemic mice with untreated NRD AML (red shaded), 72 h Dox-treated NRD AML (red open), NRI1 AML (green open) or NRI2 AML (blue open). Signaling through canonical Ras effector pathways was determined by levels of phosphorylated AKT (pAKT) for PI3K, phosphorylated ERK1/2 (pERK1/2) for MAPK and phosphorylated TBK1 (pTBK1) for RALB. Each histogram represents splenocytes from an individual mouse. All mice with NRI AMLs were maintained on Dox to prevent re-expression of NRAS(V12). The median fluorescence intensity for experimental groups is presented below the histograms for each protein of interest (error bars=1 s.d., *P<0.05, NS, nonsignificant P-value) (b) Heatmap of differential protein levels between Dox-treated de novo NRD and relapsed NRI AMLs as determined by RPPA analysis (Benjamini–Hochberg-corrected q-value of ⩽0.05).