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. 2015 Oct 8;65(1):149–163. doi: 10.2337/db15-0108

Figure 3.

Figure 3

miR-200b inhibits CITED2 expression. A: Levels of CITED2 CR, 3′-UTR, F1, or F2 (two 3′-UTR fragments encompassing the two specific binding sites of miR-200b) luciferase reporter activities after miR-200b transfection. Right: Schematic of different chimeric firefly luciferase CITED2 reporters. BS, predicted miR-200b binding site. Left: Levels of activities of luciferase reporters containing CITED2 CR, 3′-UTR, F1, or F2. Twenty-four hours after miR-200b transfection, cells were transfected with different CITED2 luciferase reporter plasmids. Levels of firefly and Renilla luciferase activities were assayed 24 h later. Results were normalized to the Renilla luciferase activities and expressed as the means ± SE data from three separate experiments (n = 3). B: Effect of deleting (Del) the two miR-200b binding sites in the CITED2 3′-UTR on luciferase reporter activities after miR-200b transfection. C: Changes in CITED2 protein expression after cells transfected with different doses of the miR-200 mimic for 48 h. D: Protein levels of CITED2 in cells transfected with the miR-200b inhibitor for 48 h. E: Levels of miR-200b in cells transfected with 75 nmol/L miR-200b for 24 h. F: Changes in CITED2 mRNA expression after miR-200b (75 nmol/L) transfection. G: miR-200b levels in cells transfected with the 75 nmol/L miR-200b inhibitor for 24 h. H: Effect of the miR-200b inhibitor (75 nmol/L) on CITED2 mRNA expression. Data were mean ± SE from three separate experiments (n = 3). *Significant differences (P < 0.05) compared with the control mimic or inhibitor groups (A, B, and E–H) or the same doses of control mimic or inhibitor groups (C and D). Ctrl, control; hRLuc, human-codon optimized Renilla luciferase.