Fig. 1

Generation and validation of miRNA-26b^−/− mice. a Schematic diagram of the generation of miR26b knockout mice. The exon-intron structure of the mouse host gene (Ctdsp1) locus is shown at the top. The targeting vector has a 2.0 kb 5′ arm including exon 2,3,4 and intron 4–5, a Neo selection cassette is flanked by FRT sites (gray bars). The loxP sites (black triangles) flanking miR26b. The 3′ recombination arm spanned 5.6 kb from the gDNA. b Southern blots show control (wild-type (1, 2, 3)) and correctly targeted heterozygote embryonic stem cell clones (4, 5,6) that gave expected hybridization patterns. c Homologous recombination was proved by primers spanning the miR26b floxed region. d Genotyping strategy based on primers identifying the wild-type, miR-26b floxed and miR-26b^−/− condition. e-g Relative mRNA expression measured by real time PCR of Ctdps1 (e), miRNA-26b-3p and miRNA-26b-5p (f), miRNA-26a-1 and miRNA-26a-2 (g). Data represent mean ± SEM, as analyzed by Mann-Whitney test