Figure 5.

Overexpression of murine miR-182 (182OE) weakens the development of MLL-AF9-transformed murine AML. (A) The representative images of BM smears by Wright-Giemsa stain in 182NC compared with 182OE AML cells (left) and statistical analysis of leukemic blasts (right). Bar scales represent 20 μm. (B) The percentages of BM GFP⁺ cells were determined in 182NC (n = 3) compared with 182OE AML cells (n = 3). Shown are the representative plots (left) and statistical analysis of GFP⁺ cells (right). (C) The percentages of BM LIC were determined in 182NC (n = 3) compared with 182OE (n = 3) AML cells. Shown are the representative plots (left) and statistical analysis of LICs (right). (D) 182NC (n = 3) and 182OE (n = 3) AML blasts were incubated with buffer containing EdU in vitro, and the incorporation of EdU was measured by flow cytometry. Shown are the representative plots (left) and statistical analysis of EdU⁺ in BM GFP⁺ cells (right). (E) BM GFP⁺ cells (2×10³/dish) from 182NC and 182OE AML mice were sorted for colony formation assay. Shown are the representative pictures (left) and statistical analysis of colony number (right). Bar scales represent 40 μm. (F and H) The representative images of the spleen (left, F) and liver (left, H) and statistical analysis of spleen weight (right, F) and liver weight (right, H). (G and I) HE stain for spleen (G) and liver (I) tissues in recipients receiving 182NC AML cells compared with those receiving 182OE AML cells at the endpoint. Bar scales represent 100 μm. (J-L) Kaplan-Meier curves and P values (log-rank test) from primary (J), secondary (K), and tertiary BMT (L) are shown, respectively. *P < 0.05; **P < 0.01; ***P < 0.001. Using log-rank test for survival time.