Fig. 5

ASPH is a direct target of miR-206 in THP-1-derived macrophages. a The overlapped eight genes in GSE54666 and GSE54039 in ox-LDL-induced THP-1 cells were shown. Among these eight genes, six genes (including ASPH) were up-regulated, while two genes were down-regulated. b The targets of miR-206 were analyzed using starbase, miRWalk and TargetScan softwares. There were 409 genes (including ASPH) that simultaneously predicted to be targets of miR-206 by these three bioinformatic softwares. c The putative binding sites between miR-206 and ASPH predicted by starbase software were shown. d and e The target relationship between miR-206 and ASPH was evaluated by dual-luciferase reporter assay and RNA-pull down assay (n = 3). The results in D were evaluated by Student’s t-test, while the results in E were assessed by ANOVA followed by Tukey’s test. f ASPH mRNA level in the serum samples of control healthy volunteers (n = 39) and AS patients (n = 39) was detected by qRT-PCR. The results were analyzed using Student’s t-test. g and h After exposing to ox-LDL for different doses or treatment times, ASPH mRNA level was measured by qRT-PCR assay (n = 3). The results were assessed by ANOVA followed by Tukey’s test. (I and J) The linear relation between the levels of ASPH mRNA and miR-206 or circTM7SF3 was analyzed by Spearman’s correlation coefficient. k and l THP-1 cells were treated with si-NC, si-circTM7SF3, si-circTM7SF3 + anti-miR-NC or si-circTM7SF3 + anti-miR-206. The mRNA and protein levels of ASPH in THP-1 cells (n = 3) were detected by qRT-PCR and Western blot assay. The protein bands shown in Fig. 5L have been cropped. The original Western blot data in Fig. 5L could be find in Additional file 2. The results were assessed by ANOVA followed by Tukey’s test. *: P < 0.05