Fig. 1

Differentiation of AML cell lines in response to 1,25D. KG1, HL60, KG1-CtrA, KG1-RARA and KG1-Ctr-len cells were exposed to 1,25D (10 or 100 nM) for 96 h and then the expression of CD11b (a) and CD14 (b) differentiation markers was detected using flow cytometry. The experiments were repeated 3–4 times and the mean percentages of positive cells (±SEM) are presented in Y-axis. The samples that differ significantly from the control are marked with asterisk (p < 0.05). To verify RARA gene silencing in KG1-RARA cells, the expression levels of RARA mRNA (c) in KG1-CtrA and KG1-RARA cell lines were measured by Real-time PCR relative to GAPDH expression levels. The bar charts show mean values (±SEM) of relative quantity (RQ). The levels of RARα protein were determined in the cytosol and nuclei of KG1, KG1-CtrA and KG1-RARA cells by western blots (d). The cytosolic (C) and nuclear (N) extracts were separated by SDS-PAGE, transferred to PVDF membranes and the proteins were revealed using anti-RARα, anti-actin and anti-HDAC antibodies