Figure 5

Dinaciclib inhibits RALB activation and has RALB-dependent antileukemic effects in human AML cell lines. (a) Western blot for RALB-GTP, total RALB and GAPDH proteins in KG1 AML cells treated for 8 h with DMSO or dinaciclib (representative of three independent experiments). (b) Phosphorylated TBK1 (pTBK1) levels in AML cells 24 h after treatment with dinaciclib (representative of three independent experiments). (c) Percentage of AML cells with cleaved PARP (cPARP+) (top) and proportion of G₀/G₁ cells (bottom) 24 h after treatment with dinaciclib determined by flow cytometry (n=3 independent experiments, error bars=s.e.m., *P<0.05). (d) MTS viability analysis of AML cell lines 72 h after dinaciclib treatment and calculated half-maximal inhibitory concentrations (IC₅₀) (n=3–5 independent experiments, error bars are not included for clarity of presentation and are included in Supplementary Figure 4). (e) Relative leukemic colony formation (L-CFC) of THP1 transduced with RALB(Q72L), myristoylated AKT (myrAKT), CRAF 22W or control vector (BFP) 24 h after treatment with dinaciclib relative to DMSO-treated controls (n=3 independent experiments, error bars=1 s.d., *P<0.05).