Fig. 5

MiR-182 blocks granulocytic differentiation in vitro. a Overexpression of miR-182 reduced the amount of C/EBPα protein in both DMSO treated and ATRA treated U937 cells as analyzed by western blot. b and c Enforced expression of miR-182 led to a reduced percentage of CD11b-positive U937 cells after treatment with ATRA. b Representative FACS analysis for myeloid marker CD11b in miR-182 or scramble transfected U937 cells treated for 24 or 48 h with 1 µM ATRA to induce granulocytic differentiation and c summary of all experimental repetitions. Data represent the mean ± SD of three independent experiments. (*p < 0.05) P values were calculated using unpaired Student’s t-test. d Overexpression of miR-182 did not affect the amount of C/EBPα-ER fusionprotein in K562-C/EBPα-p42-ER cells as analyzed by western blot. e and f Enforced expression of miR-182 did not affect the percentage of CD11b-positive K562-C/EBPα-ER cells after treatment with β-estradiol. e Representative FACS analysis for myeloid marker CD11b in miR-182 or scramble transfected K562-C/EBPα-ER cells treated for 24 or 48 h with 5 µM β-estradiol to induce granulocytic differentiation and f summary of all experimental repetitions. Data represent the mean ± SD of three independent experiments. g Block of miR-182 by decoy lentiviral infection increased the amount of C/EBPα protein level in Kasumi-1 cells as analyzed by western blot. h and i A block of miR-182 accelerates myeloid differentiation of Kasumi-1 cells induced by 1 mM Sodium phenylbutyrate (SPB). h Representative FACS analysis for myeloid marker CD11b in decoy-miR-182 or control infected Kasumi-1 cells treated for 72 h with either vehicle control (left panel) or 1 mM SPB to induce granulocytic differentiation and i summary of all experimental repetitions. Data represent the mean ± SD of four independent experiments. (*p < 0.05) P values were calculated using unpaired Student’s t-test