Figure 2

CTGF is targeted by miR-26a and miR-30c, and Snail1 is targeted by miR-30c.

(A) Sequences of the CTGF 3′-UTR containing the miR-26a and miR-30c binding sites and the Snail1 3′-UTR containing the miR-30c binding site. The mutant sequences were used in the mutant 3′-UTR constructs. (B,C) NRK-52E cells were co-transfected with CTGF or Snail1 3′-UTR luciferase reporter plasmids (1 μg), β-galactosidase plasmid (0.2 μg), and miR-26a or miR-30c mimic alone (50 nM) or in combination (25 nM each) following the various indicated treatments. The luciferase and β-gal activities were detected using a luciferase assay kit after 72 h. The bars represent the mean ± SEM (n = 3). *p < 0.05 versus NC; ^#p < 0.05 versus miR-26a mimic; ^$p < 0.05 versus miR-30c mimic. (D,E) qRT-PCR analysis and (F) western blots of CTGF and Snail1 in NRK-52E cells transfected with miR-26a and miR-30c mimics for 72 h. The bars represent the mean ± SEM (n = 3). *p < 0.05 versus NC; ^#p < 0.05 versus TGFβ1; ^$p < 0.05 versus miR-26a mimic+TGFβ1; ^&p < 0.05 versus miR-30c mimic+TGFβ1.