Figure 6.

MFSD4A-AS1 sponges miR-30c-2-3p, miR-145-3p and miR-139-5p. (A) Volcano plot analyzed the clinical correlation of MFSD4A-AS1 with all reported miRNAs in the PTC dataset from TCGA. The orange colors represent significantly and negatively correlated miRNAs with fold change > 2 and r value < −0.2. (B) Real-time PCR analysis of the effect of MFSD4A-AS1 on expression of miR-30a-5p, miR-126-3p, miR-126-5p, miR-30c-2-3p, miR-145-3p, miR-30c-5p, miR-152-3p, miR-30a-3p, miR-139-5p and miR-139-3p. Pseudo-color scale values were log₂ transformed. Transcript levels were normalized by U6 expression. The experiment was independently performed three times. (C and D) The effect of miR-30c-2-3p, miR-145-3p and miR-139-5p on the luciferase activity of wild-type or mutant MFSD4A-AS1 in the indicated cells. Error bars represent the mean ± s.d. of three independent experiments. *P < 0.05. (E and F) RNA immunoprecipitation (RIP) assay showing the association among miR-30c-2-3p, miR-145-3p and miR-139-5p, and MFSD4A-AS1 transcripts in PTC cells. Pulldown of IgG antibody served as the negative control *P < 0.05. (G) The mediating effect of changed expression of miR-139-5p, miR-145-3p and miR-30c-2-3p in PTC cells on human umbilical vein endothelial cells (HUVECs) in tube formation assay. Each bar represents the mean values ± s.d. of three independent experiments. *P < 0.05. (H) The mediating effect of changed expression of miR-139-5p, miR-145-3p and miR-30c-2-3p in PTC cells on migration ability of HUVECs in the Transwell assay. Each bar represents the mean values ± s.d. of three independent experiments. *P < 0.05.