Figure 1

MiR-10a-induced inhibition of hoxd4 gene expression. (a) Schematic representation of the miR-10a&b loci upstream of the hoxd4 and hoxb4 genes. The insert shows the ncRNA loci, the BSP analysed region, the siRNA target sites, and the regions analysed by ChIP in the Hoxd4 promotor. (b) MiRNA10 exhibits high evolutionary conservation in human, mouse, zebrafish, Drosophila and chicken. (c) Profile of miR-10a/b expression in cancer cells by real time PCR (1: MCF7; 2: MDA-MB-231; 3: MCF10A: 4: HepG2; 5: HeLa; 6: A549). (d) Quantitative RT-PCR analysis of hoxd4 and hoxb4 gene expression in human cancer cell lines (1: MCF7; 2: MDA-MB-231; 3: MCF10A: 4: HepG2; 5: HeLa; 6: A549). The expression miR-10a varies inversely with the expression of hoxd4 in all six cell types. (e) Quantitative PCR analysis of the hoxd4 RNA levels in MCF7 and MDA-MB-231 cells treated with antisense miR-10a 2'-O-methyl oligos, miR-196a 2'-O-methyl oligos or negative control oligos (N.C.). (f) RT-PCR analysis of the hoxd4, hoxd3 and hoxd8 mRNA levels in MCF7 and MDA-MB-231 cells treated with antisense miR-10a 2'-O-methyl oligos or negative control oligos (N.C.). (g) Protein analysis of Hoxd4 in MDA-MB-231 cells after transfection with antisense miR-10a 2'-O-methyl oligos, miR-196a 2'-O-methyl oligos or negative control oligos (N.C.).