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. Author manuscript; available in PMC: 2012 Jan 1.
Published in final edited form as: Inflamm Bowel Dis. 2011 Jan;17(1):221–231. doi: 10.1002/ibd.21359

Figure 5. A.Interaction of miR-31 with FIH1 3’UTR.

Figure 5

Figure 5

Figure 5

HCT-116 colon cancer cells were transfected with miR-31 mimic or a non-specific control mimic (NSM). After 24h, cells were co-transfected with a construct containing a truncated FIH1 3’UTR containing two miR-31 putative binding sites fused to the firefly luciferase gene. As a negative control, the 3’UTR was cloned in reverse orientation. When FIH1 3’UTR forward orientation was used, a moderate but statistically significant difference was observed in miR-31-transfected cells compared to the nonspecific mimic. This inhibitory effect disappeared with inversion of the FIH1 3’UTR. B. Effect of miR-31 on FIH1 protein levels. Western blotting was performed on cells transfected with miR-31. As a negative control, cells were transfected with either 1) a non-specific mimic, 2) miR-21 (which has no predicted binding site in the 3’UTR of FIH1), or 3) miR-224 (which does have one predicted binding site within the FIH1 3’UTR). C. Quantification of FIH1 protein levels. Densitometry of Western blots was carried out using Image J software.