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. 2019 Feb 27;58(6):1019–1032. doi: 10.1002/mc.22990

Figure 4.

Figure 4

NRP1 is a direct target gene of miR‐338‐3p, an miRNA whose expression is decreased in NSCLC tissues and cell lines. A, Schematic diagram showing the subcloning of the predicted miR‐338‐3p binding site at two positions (234‐240 and 353‐359) in the NRP1 3′‐UTR into the psiCHECK‐2 luciferase vector. The predicted duplex formation between miR‐338‐3p and the wild‐type or mutant miR‐338‐3p binding site on NRP1 is indicated. B, Luciferase activity of the construct containing the wild‐type or mutant NRP1 3′‐UTR reporter gene in A549 and H226 cells co‐transfected with negative control (NC) or miR‐338‐3p. Scrambled sequences were used as the NC. The relative Renilla luciferase activity was determined and normalized to the firefly luciferase activity. C‐E, Expression of miR‐338‐3p and NRP1 in NSCLC cells transfected with the miR‐338‐3p mimics or inhibitor was detected by qRT‐PCR and Western blotting, respectively. F, Relative miR‐338‐3p levels in 55 NSCLC tissues (T) and paired noncancerous lung tissues (N). G, Scatter plot showing the relative miR‐338‐3p expression levels in NSCLC tumor and adjacent normal lung tissues from a public dataset (GSE36681). H, QRT‐PCR analysis of relative miR‐338‐3p expression in human NSCLC cell lines and a human bronchial epithelial cell line. [Color figure can be viewed at wileyonlinelibrary.com]