Figure 7.

LPAR5 is the target gene for miR-513c-5p. (A) The putative binding site of LPAR5 and miR-513c-5p is shown. (B) Luciferase activity was measured in SW579 and TPC-1 cells co-transfected with wild type (LPAR5-Wt) or mutant (LPAR5-Mut) LPAR5 and miR-513c-5p using the luciferase reporter assay. **P < 0.001 vs miR-NC. (C) Biotinylated RNA pull-down assays were applied using Bio-NC and Bio-mimic. Relative enrichment of LPAR5 was measured by qRT-PCR. Bio-mimic, miR-513c-5p mimic. **P < 0.001 vs Bio-NC. (D) The expression level of LPAR5 was detected by qRT-PCR in THCA tissues and paired adjacent normal tissues. (E) Pearson correlation analysis of the correlation between LPAR5 and miR-513c-5p in THCA tissues. (F) The expression level of LPAR5 was measured by qRT-PCR in the human thyroid follicular epithelial cell line (Nthy-ori 3–1) and THCA cell lines (TPC-1 and SW579). **P < 0.001 vs Nthy-ori 3–1. (G) The mRNA expression level of LPAR5 were measured by Western blot in SW579 and TPC-1 cells transfected with miR-513c-5p mimic or miR-513c-5p inhibitor. Inhibitor, miR-513c-5p inhibitor. Mimic, miR-513c-5p mimic. (H) The protein expression level of LPAR5 were measured by Western blot in SW579 and TPC-1 cells transfected with miR-513c-5p mimic or miR-513c-5p inhibitor. Mimic, miR-513c-5p mimic. Inhibitor, miR-513c-5p inhibitor. Mimic, miR-513c-5p mimic. **P < 0.001 vs Mimic-NC; ## P < 0.001 vs Inhibitor-NC.

Abbreviations: Bio-NC, Biotin labeled negative control of miR-513c-5p mimic.