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. 2008 Jan 14;99(2):280–286. doi: 10.1111/j.1349-7006.2007.00666.x

Figure 4.

Figure 4

Induction of miR‐138 in HEK‐293 cells. (a) Western blotting for human telomerase reverse transcriptase (hTERT) protein. Nuclear extract (30 µg) was applied to each lane. Equal loading was confirmed by blotting of lamin‐B. hTERT protein expression was decreased in HEK‐293 transfected with miR‐138 precursor molecules in comparison with negative control precursor miRNA (N/C). (b) Amplification plots of hTERT mRNA in HEK‐293 transfected with miR‐138 precursor molecules. There was no difference in hTERT mRNA expression before and after transfection. Relative hTERT mRNA expression data were obtained using the ρρcycle threshold (Ct) method with glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) as an endogenous control. (c) Base pairing for comparison between mature miR‐138 and the wild‐type (WT) or mutant (MUT) putative target sites in the 3′‐untranslated region (UTR) of hTERT mRNA. (d) Luciferase assay of HEK‐293 transfected with luciferase constructs containing the WT (clear bar) or MUT (solid bar) target site of the 3′‐UTR of hTERT mRNA. Values represent mean ± SE of six experiments from three independent transfections.