Research Paper Volume 15, Issue 24 pp 15624—15639

A regulatory circuit of lncRNA NLGN1-AS1 and Wnt signalling controls clear cell renal cell carcinoma phenotypes through FZD4-modulated pathways

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Figure 6. FZD4 is a direct target of miR-136-5p in ccRCC progression. (A) Relative expression of FZD4 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. (B) Relative expression of FZD4 in 769-P and Caki-1 cells transfected with miR-136-5p mimic. (C) Western blots analysis of FZD4 in miR-136-5p mimic 769-P and Caki-1 cells compared with NC group, GAPDH was used as a loading control. (D) Schematic illustration of the predicted binding sites between FZD4 and miR-136-5p, and mutation of potential miR-136-5p binding sequence in FZD4. Relative luciferase activities of wild type (WT) and mutated (Mut) FZD4 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.