Fig. 3

CircTM7SF3 directly interacts with miR-206 in THP-1-derived macrophages. a The miRNA targets of circTM7SF3 were predicted by starbase software. Among these putative miRNA targets, five miRNAs (miR-320b, miR-613, miR-425-5p, miR-206 and miR-34a-5p) have been reported to be down-regulated in AS. b The expression of miR-320b, miR-613, miR-425-5p, miR-206 and miR-34a-5p in differentiated THP-1 cells transfected with si-NC or si-circTM7SF3 was measured by qRT-PCR (n = 3). The results were analyzed using Student’s t-test. c The target sequence of miR-206 in circTM7SF3 that predicted by starbase software was shown. Also, the mutant binding sites with miR-206 in circTM7SF3 were also shown. d Dual-luciferase reporter assay was performed to verify the interaction between miR-206 and circTM7SF3 in THP-1 cells. THP-1 cells (n = 3) were co-transfected with miR-NC or miR-206 and circTM7SF3 wild-type reporter plasmid (circTM7SF3-WT) or circTM7SF3 mutant reporter plasmid (circTM7SF3-MUT). The results were analyzed using Student’s t-test. e RNA-pull down assay was carried out to test the target interaction between miR-206 and circTM7SF3 in THP-1 cells (n = 3). The results were assessed by ANOVA followed by Tukey’s test. f MiR-206 level in the serum of Control group (n = 39) and AS patients (n = 39) was analyzed by qRT-PCR. The results were analyzed using Student’s t-test. g The linear correlation between the expression of circTM7SF3 and miR-206 in the serum of AS patients (n = 39) was analyzed using Spearman’s correlation coefficient. h and i MiR-206 level in THP-1 cells (n = 3) treated with different doses or times of ox-LDL was analyzed by qRT-PCR. The results were assessed by ANOVA followed by Tukey’s test. j MiR-206 level was examined in THP-1 cells (n = 3) transfected with si-NC or si-circTM7SF3 by qRT-PCR. The results were analyzed using Student’s t-test. *: P < 0.05