Fig. 8

Enforced expression of miR-182 blocks granulocytic differentiation in vivo. a Schematic overview of the LSK based transplantation model. b MiR-182 levels in GFP⁺ LSK cells transduced with either miR-182 or scramble lentiviral particles. MiR-182 expression was measured by qPCR 48 h after transduction. c Representative flow cytometry analysis from blood (4 and 10 weeks) and bone marrow (10 weeks) after transplantation revealed a reduced percentage of Mac1⁺/Gr1⁺ granulocytes in the GFP⁺ fraction of mice transplanted with miR-182 transduced LSK cells compared to mice transplanted with scramble transduced cells. GFP⁻ cells served as controls. d Summary of flow cytometry analysis from blood and bone marrow at indicated time points after transplantation. Percentage of mature Mac1⁺/Gr1⁺ granulocytes in the GFP⁺ fraction of mice transplanted with miR-182 transduced LSK cells was significantly reduced. No effect was observed in the GFP⁻ fraction. Bars represent the summary of 3 independent bone marrow transplantations with the indicated total number of animals (4 weeks: scramble n = 9 and miR-182 n = 8; 10 weeks: scramble n = 8 and miR-182 n = 6; *p < 0.05; **p < 0.01; ns—not significant). P values were calculated using unpaired Student’s t-test. e MiR-182 expressing mice showed reduced CEBPA mRNA levels in the bone marrow. GFP + cells from the bone marrow 10 weeks after transplantation were sorted and analyzed by qPCR for CEBPA mRNA expression. Data represent the mean ± SD of four independent animals. (*p < 0.05) P values were calculated using unpaired Student’s t-test. f and g Cell morphology analysis from GFP⁺ sorted bone marrow cells 10 weeks after transplantation by Wright-Giemsa-staining showed reduced numbers of mature neutrophils in miR-182 transduced mice. f Representative photographs (Scale bar 5 µM) and g summary of four mice (scramble) and three mice (miR-182), respectively. (*p < 0.05) P values were calculated using unpaired Student’s t-test