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. 2015 Jul 1;8(7):8048–8056.

Figure 3.

Figure 3

MiR-192 directly inhibits the expression of Bim through its 3’UTR of ESCC cells. A. The miR-192 binding site predicted in the 3’UTR of Bim mRNA. B. Mutant was generated at the seed region of Bim 3’UTR as indicated by the underline. A 3’UTR fragment of Bim mRNA containing wild-type or mutant of the miR-192 binding sequence was cloned into the downstream of the luciferase gene in pMIR vector. C. KYSE-150 cells was transfected with pMIR reporter vectors containing either wild-type or mutant Bim 3’UTR (indicated as pMIR-Bim-3’UTR-wt and pMIR- Bim-3’UTR-mut) with either pcDNA3.1 (indicated as NC) or pcDNA3.1-miR-192 vector (indicated as miR-192). Luciferase activity was determined 48 h after transfection. D. Bim mRNA was detected by qRT-PCR in cell lines transfected with pcDNA3.1 (indicated as NC) or pcDNA3.1-miR-192 vector (indicated as miR-192). E, F. The protein levels of Bim was examined by Western blot in cells transfected with different plasmids. Figure F shows the relative gray values of each band (normalized to β-actin). Data are reported as mean ± SD (*P < 0.01, Student’s t- test).