Fig. 4: MiR-21 targets KBTBD7.

a Shown is the alignment of miR-21, its target sites and mutant in 3′-UTR of KBTBD7 (left). Luciferase activity assay in lysates of HEK293 cells 24 h after transfection with wild-type or mutant KBTBD7 3′-UTR firefly luciferase reporter plasmids, together with miR-21 or control mimics. The firefly luciferase activity was normalized by renilla luciferase activity (Right). Data represent the mean ± SEM (n = 6 independent preparations). **P < 0.01 vs. ctrl mimics (two-way ANOVA). b Q-PCR analysis of Kbtbd7 mRNA (normalization to GAPDH) or immunoblot analysis of its protein expression in heart tissues of remote area (RA) or border area (BA) from miR-21 KO or WT mice 3 days after MI. Data represent the mean ± SEM (n = 6 mice per group). **P < 0.01 vs. WT-MI-RA (two-way ANOVA). c Q-PCR analysis of Kbtbd7 mRNA (normalization to GAPDH) or immunoblot analysis of its protein expression in heart tissues of remote area (RA) or border area (BA) from mice 3 days after MI followed by injection with miR-21 (Ad-miR-21) or control adenovirus (Ad) into peri-infarct heart tissue. Data represent the mean ± SEM (n = 6 mice per group). **P < 0.01 vs. MI-Ad-BA (two-way ANOVA). d Q-PCR analysis of Kbtbd7 mRNA (normalization to β-actin) or immunoblot analysis of its protein expression in miR-21-deficient or WT macrophages. Data represent the mean ± SEM (n = 6 independent preparations). **P < 0.01 vs. WT (Student’s t-test). e Immunoblot analysis of KBTBD7 protein expression in macrophages 48 h after transfection with miR-21 or control mimics