Research Paper Volume 12, Issue 7 pp 5948—5976

Figure 11. HOXA3 acts as a targeted gene of miR-10a-5p. (A) Volcano plot of differentially expressed mRNAs between control group and IL-1β induced primary chondrocyte group from GSE74220. (B) Venn plot of the overlapping mRNA across the downregulated mRNAs in GSE74220, the predicted targets of miR-10a-5p in Targetscan, and the downregulated mRNAs in our RNA-seq. (C) The expression level of HOXA3 in OA cartilage from GSE114007, **P < 0.01. (D) Targetscan shows that the seed region of miR-10a-5p is well-paired with 3’UTR of HOXA3, which was conservative among vertebrates. (E) High-throughput experiments verified that HOXA3 was the direct target of miR-10a-5p in HEK293, TZMBL, and 293S from TarBase v.8. (F) The sequencing results of cloned fragments in luciferase reporter vectors and the yellow highlighted sequence is the target test site. (G) Interaction between miR-10a-5p and HOXA3 was verified by luciferase report assay in 293T cells. (H, I) The relative expression of HOXA3 in control and IL-1β induced Hc-a analyzed by RT-qPCR and western blot (n=3, *P < 0.05). (J, K) The relative expression of HOXA3 after transfecting miR-10a-5p mimics analyzed by RT-qPCR and western blot (n=3, **P < 0.01).