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. 2018 Jan 18;41(4):1855–1866. doi: 10.3892/ijmm.2018.3405

Figure 5.

Figure 5

PTEN and TIMP3 are targets of miR-21-5p. (A) The 3′-UTRs of PTEN and TIMP3 were identified as putative targets of miR-21-5p with 7-mer or 8-mer complementarity regions. Seven nucleotides in the miR-21-5p seed region (gray font) were mutated using polymerase chain reaction site-directed mutagenesis. (B) A dual-luciferase reporter assay was performed to assess the influence of miR-21-5p on luciferase intensity controlled by the 3′-UTR of potential targets. (C and D) Western blot analysis indicated a decrease or increase in PTEN and TIMP3 protein levels following transfection of miR-21 mimics or anti-miR-21, respectively. Western blot images are representative of at least three independent experiments. The value under each sample indicates the fold change of PTEN or TIMP3 protein levels relative to the control. *P<0.05 vs. miR-NC. PTEN, phosphatase and tensin homologue; TIMP3, tissue inhibitor of matrix metalloproteinases 3; UTR, untranslated region; miR, microRNA; NC, negative control; Hsa, Homo sapiens; MUT, mutant; SGC7901/DOX, doxorubicin-resistant SGC7901 cell line.