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. 2016 Feb 20;7(12):14569–14585. doi: 10.18632/oncotarget.7538

Figure 1. MiR-410 directly targeted SLC34A2.

Figure 1

A. Four miRNAs (miR-410, miR-491-5P, miR-384 and miR-506-3P) were predicted by both algorithms (TargetScan, miRanda). The numbers indicate the nucleotide position in reference to the start of SLC34A2 3′UTR. B. The expression of miR-410, miR-491-5P, miR-384 and miR-506-3P in A549 cells was determined by qRT-PCR. C. The expressions of miR-410 in A549, 95D and H1299 cells were determined by qRT-PCR. D. Relative expression of miR-410 and SLC34A2 detected by qRT-PCR in NSCLC patient tissues. Increased miR-410 expression and decreased SLC34A2 expression were indicated in 9 of 12 NSCLC patient tissues compared with adjacent non-tumorous tissues. E. Luciferase reporter assay was performed to confirm the miR-410 binding to the 3′UTR of SLC34A2. The luciferase activity was detected after co-transfection with luciferase reporter plasmids (Pmir-SLC34A2 3′UTR-F, P-SLC34A2-F; Pmir-SLC34A2 3′UTR-R, P-SLC34A2-R), with miR-410 mimics/NC or miR-410 inhibitors/NC in HEK293 cells. F. Real-time PCR was performed to detect SLC34A2 mRNA level after transfection of miR-410 inhibitors or miR-410 mimics with corresponding control in A549 cells. G. Western blotting was performed to detect SLC34A2 protein level after transfection of miR-410 inhibitors with corresponding control in A549 cells. For miR-410 and SLC34A2 mRNA expression detected by qRT-PCR, U6 and β-actin were used as internal control respectively. For SLC34A2 protein expression detected by western blotting, β-actin was used as internal loading control. Data are presented as the mean value ± SD from triplicate experiments. *, p < 0.05; **, p < 0.01.