Figure 5.

Target Prediction and Verification of Their Regulation via Endoderm- and Mesoderm-Specific miRNAs

(A) Predicted targets of miR-1263 and miR-483-3p calculated by the TargetScan or miRanda algorithm.

(B) Tabular presentation of the candidate genes predicted by both algorithms for miR-1263 and miR-483-3p. Names written in bold were experimentally analyzed by luciferase reporter assays.

(C) Luciferase activity normalized to the β-galactosidase activity for the subcloned 3′ UTRs of KLF4, PGAM1, and ZBTB26 in HEK293 cells upon transfection of the indicated mimics and the reporter plasmids. Values are mean ± SEM, n = 4–6. Two-tailed Student's t test, ^∗∗p < 0.01.

(D and E) Normalized protein expression of KLF4 (D) and PGAM1 (E) 24 hr and 48 hr post transfection with the indicated miRNA mimics in HEK293. The NC of each experiment was set as 100%. For each protein one representative western blots is shown on the right. Values are mean ± SEM, n = 4–5. ANOVA plus Bonferroni's post hoc test, ^∗p < 0.05 and ^∗∗p < 0.01 compared with the respective NC.

See also Figure S5.