Figure 2
Effect of TAK165 on ATRA-induced AML cell differentiation.
(a) HL60 and NB4 cell proliferation assay. The cells were treated with the indicated concentrations of TAK165 in the presence of vehicle or ATRA (1 μM for HL60, 2 nM for NB4) for 3 days. (b) HL60 cell flow cytometric cycle proportion assay. The cells were treated with 100 nM TAK165 in the presence of vehicle or 1 μM ATRA for 3 days. (c) CD11b expression in the HL60 and NB4 cells. The cells were treated with the indicated concentrations of TAK165 in the presence of vehicle or ATRA (1 μM for HL60, 2 nM for NB4) for 3 days. (d) NB4 cell NBT-reducing activity. The cells were treated with 50 nM TAK165 in the presence of vehicle or 2 nM ATRA for 3 days. (e) HL60 and NB4 cell morphologic differentiation. The cells were treated with the indicated concentrations of TAK165 in the presence of vehicle or ATRA (1 μM for HL60, 2 nM for NB4) for 3 days. (f) CEBPB, CEBPE and PU.1 mRNA levels in the HL60 cells as determined by real-time PCR. The cells were treated with 100 nM TAK165 in the presence of 1 μM ATRA for 1 day. GAPDH expression was used as an internal control gene. (g) Western blot analysis of PU.1 and C/EBPβ in the HL60 cells. The cells were treated with 100 nM TAK165 in the presence of 1 μM ATRA for 3 day. In (a,c,f), the data are presented as the mean ± SD of 3 independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001.
