Research Paper Volume 15, Issue 24 pp 15624—15639

Figure 4. NLGN1-AS1 binds to miR-136-5p and represses its expression. (A) A schematic diagram used to search the target miRNAs of NLGN1-AS1 in three databases. (B) qRT-PCR assay confirmed the relative expression of five candidate miRNAs of NLGN1-AS1 in 769-P and Caki-1 cells and 20 paired ccRCC cancer tissues compared with corresponding adjacent normal tissues. (C) Relative expression of miR-136-5p in a series of RCC cell lines (786-O, 769-P, Caki-1, Caki-2, ACHN, A498 and OSRC-2) and human normal renal tubular epithelial cell line HK-2. (D, E) Relative expression of miR-136-5p in 769-P and Caki-1 cells transfected with miR-136-5p mimic(d) or cells after transfection with sh-NLGN1-AS1(e). (F) Relative expression of NLGN1-AS1 in 769-P and Caki-1 cells transfected with miR-136-5p mimic. (G) Schematic illustration of the predicted binding sites between NLGN1-AS1 and miR-136-5p, and mutation of potential miR-136-5p binding sequence in NLGN1-AS1. Relative luciferase activities of wild type (WT) and mutated (Mut) NLGN1-AS1 reporter plasmid in human embryonic kidney (HEK) 293T cells co-transfected with miR-136-5p mimic. The data represent the mean ± SD of 3 replicates. ^*P < 0.05; ^**P < 0.01.