Fig. 4: miR-141-3p inhibits FUS expression in NP cells.

a The Venn diagram indicates the 19 predicted miRNAs regulate FUS expression. miR-141-3p was intersected predicted by 5 different databases. b Expression of miR-141-3p in IDD NP tissues, showing that miR-141-3p expression was significantly higher than that of controls. Quantitative data from three independent experiments is presented as mean ± SEM (error bars). ***P < 0.001. c Sequence alignment of a putative miR-141-3p-binding site within the 3′UTR of FUS mRNA. Bottom: mutations in the 3′UTR of FUS mRNA sequence to create the mutant luciferase reporter constructs. d Luciferase reporter assay in NP cells after transfected with scramble oligo or miR-141-3p mimics, Renilla luciferase vector, and the reporter constructs. Both firefly and Renilla luciferase activities are measured in the same sample. Firefly luciferase signals were normalized with Renilla luciferase signals. Quantitative data from three independent experiments is presented as mean ± SEM (error bars). ***P < 0.001. e, f FUS expression level was detected by qRT-PCR, western blot in primary human NP cells. Three independent experiments is presented as mean ± SEM (error bars). ***P < 0.001. g NP cells from control tissues were transfected with miR-141-3p mimic or miR-141-3p inhibitor. qRT-PCR was used to detect the relative expression level of circ-GRB10 compared with controls. Three independent experiments is presented as mean ± SEM (error bars). ***P < 0.001. h NP cells from control tissues were transfected with miR-141-3p with or without FUS overexpress plasmid. qRT-PCR was used to detect the relative expression level of circ-GRB10 compared with controls. i miR-141-3p inhibitor with or without FUS siRNA was transfected into NP cells from control tissues and the expression level of FUS. Three independent experiments is presented as mean ± SEM (error bars). ***P < 0.001.