Figure 5. MiR-320d targets E2F1 in NSCLC cells. (A) The interaction of miR-320d and E2F1 3’ UTR was identified by bioinformatic analysis using Targetscan (http://www.targetscan.org/vert_72/). (B–E) The HCC827 and A549 cells were treated with miR-320d mimic or the control mimic. (B) The expression of miR-320d was tested by qPCR assays in the cells. (C) The luciferase activities of wild type E2F1 (E2F1 WT) and E2F1 with the miR-320d-binding site mutant (E2F1 MUT) were determined by luciferase reporter gene assays in the cells. (D) The mRNA expression of E2F1 was measured by qPCR assays in the cells. (E) The protein expression of E2F1 was analyzed by Western blot analysis in the cells. (F) The HCC827 and A549 cells were treated with the control vector, LINC00662 overexpression vector, or co-treated with LINC00662 overexpression vector and miR-320d mimic. The protein expression of E2F1 was measured by Western blot analysis in the cells. Data are presented as mean ± SD. Statistic significant differences were indicated: * P < 0.05, ** P < 0.01.