ZEB2-targeting miR-200 family miRNAs are epigenetically silenced in AML. (A) qPCR of ZEB2, miR-200b, and miR-200c in a panel of AML cell lines and epithelial cancer cell line MCF-7. (B) Expression of miR-200 family members and ZEB2 in cancer samples from TCGA RNA-Seq data; expression level is shown as log2 of reads per kilobase of transcript per million mapped reads. (C) Methylation of DNA sequences upstream of miR-200c and miR-141 in TCGA AML samples. Red arrows indicate the relative position of miR-200c and miR-141. Each dot indicates average level of methylation in all patients. (D) miR-200c is upregulated after 3 days of 5-aza treatment as measured by qPCR. (E) Viability of AML cell lines after miR-200c and empty vector infection. Cells were seeded at day 6 after infection, and cell viability was measured at day 9 by the CellTiter-Glo assay. (F) Immunoblot of ZEB2 in cells infected with miR-200c. (G) Competitive proliferation assay in mouse leukemia cells infected with mouse miR-200c or control vector. Error bars, SD. n = 190 for AML, n = 4837 for epithelial cancers in B; n = 193 for C; and n = 3 for D and E.
