Fig 1: MAPK8IP1P2 activates Hippo signaling by sponging miR-146b-3p. (A) Volcano plot analyzed the clinical correlation of MAPK8IP1P2 with all reported miRNAs in thyroid cancer dataset from TCGA. The orange colors represent significantly and negatively correlated miRNAs with fold change > 2 and r value < -0.2. (B) Predicted recognition sites of miR-146b-3p on MAPK8IP1P2, and predicted miR-146b-3p targeting sequence and mutant sequences in 3’UTR s of NF2, RASSF1, and RASSF5. (C, D) The effect of miR-146b-3p on the luciferase activity of wild-type or mutant MAPK8IP1P2, NF2, RASSF1 and RASSF5 in the indicated cells. Error bars represent the mean ± S.D. of three independent experiments. *P < 0.05. (E, F) Real-time PCR analysis of the effect of overexpression or silencing MAPK8IP1P2 on NF2, RASSF1, and RASSF5 expression in the indicated cells. Transcript levels were normalized by GAPDH expression. Error bars represent the mean ± S.D. of three independent experiments. *P < 0.05. (G) Western blot analysis of the effect of overexpression or silencing MAPK8IP1P2 on NF2, RASSF1 and RASSF5 expression in the indicated cells. α-Tubulin served as the loading control. (H, I) Real-time PCR (H) and Western blot (I) analysis of the effect of miR-146b-3p mimics on NF2, RASSF1, and RASSF5 expression in MAPK8IP1P2-overexpressing thyroid cancer cells. Transcript levels were normalized by GAPDH expression. α-Tubulin served as the loading control. Error bars represent the mean ± S.D. of three independent experiments. *P < 0.05. (J) The effect of miR-146b-3p mimics on TEAD transcriptional activity was assessed by HOP-Flash luciferase reporter in MAPK8IP1P2-overexpressing thyroid cancer cells. Error bars represent the mean ± S.D. of three independent experiments. *P < 0.05.