Research Paper Volume 14, Issue 24 pp 10009—10026

Figure 3. circ_0066715 acts as a miRNA sponge-like effect of miR-486-5p. (A) Nucleocytoplasmic separation experiments in RA-FLS cells. (n=9). (B) circ_0066715 localization in RA-FLS cells.(IF,×200). (C) Heatmap of differentially expressed miRNAs in PBMC of RA patients. (n=3). (D) Volcano plot of differentially expressed miRNAs in PBMC of RA patients. (n=3). (E) Circ_0066715 binding map with miRNA targeting. (F) RNA_ pull-down assay enrichment result of circ_0066715.**P < 0.01, vs Negative. (n=3, paired t-test). (G). RNA pull-down assay enriched miR-486-5p results. **P < 0.01, vs Negative. (n=3, paired t-test). (H) Binding site between miR-486-5p and circ_0066715. (I) Luciferase reporter assay. * P < 0.05, vs hsa_circ_0066715-Mut. (n=3, paired t-test). (J) Analysis of cell viability after targeting miR-486-5p with circ_0066715.**P < 0.01. (n=3, ANOVA with Bonferroni’s post test). (K) Clonal analysis of cells after targeting miR-486-5p with circ_0066715. **P < 0.01. (n=3, ANOVA with Bonferroni’s post test). (L) Cytokine expression after circ_0066715 targeted miR-486-5p in RA patients. * P < 0.05, vs RA-FLS+pcDNA3.1-circ_0066715. (n=15, paired t-test).