Figure 3 | Scientific Reports

Figure 3

From: MicroRNA-130b functions as an oncomiRNA in non-small cell lung cancer by targeting tissue inhibitor of metalloproteinase-2

Figure 3

miR-130b targeted TIMP-2 in NSCLC cells. (A) A549 cells stably overexpressing miR-130b were transfected with a luciferase reporter construct containing the predicted miR-130b-binding site (WT) or the mutated predicted miR-130b-binding site (mut) in the TIMP-2 3′-UTR. Data are means ± standard deviations of more than five independent experiments. *p < 0.05 for one-way analysis of variance followed by post-hoc Bonferroni multiple comparison tests. (B) A549 cells stably overexpressing miR-130b (130b) or control vector (mock) were subjected to real-time qPCR analysis of TIMP-2. Relative expression of TIMP-2 normalized to GAPDH is shown (means ± standard deviations) from three independent experiments. (C,D) A549 cells stably overexpressing miR-130b (130b) or control vector (mock) were subjected to western blotting with anti-TIMP-2 antibodies. Representative results from three independent experiments are shown. Representative results of three independent experiments are shown for (C). The numbers indicate the relative expression of TIMP-2 compared to that in NC, as analyzed by densitometry. *p < 0.05 for t-tests. Uncropped western blot data is shown in Supplementary Fig. 10. (E,F) NCI-H1755 cells were transfected with the negative control inhibitor (NC) or miR-130b inhibitor (130b) for 48 h, and whole-cell lysates were subjected to western blotting with anti-TIMP-2 antibodies. Representative results of three independent experiments are shown. The numbers indicate the relative expression of TIMP-2 compared to that in NC, as analysed by densitometry. *p < 0.05 for t-tests. Uncropped western blot data is shown in Supplementary Fig. 10.

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