Research Paper Volume 16, Issue 4 pp 3896—3914

Figure 2. LncRNA GAS5 is upregulated in IVDD-derived AFs and inhibits miR-221-3p expression. (A) qRT-PCR was adopted to examine the expression of miR-221-3p in AFs. (n = 6) (B) The Venn diagram was drawn to obtain the common lncRNA of upregulated lncRNA in IVDD based on the GSE56081 dataset, and ENCORI predicted the potential target lncRNA of miR-221-3p. (C) The expression level of GAS5 between normal and degenerative disks on the GSE56081 dataset. (n = 5) (D) qRT-PCR assay was utilized to detect the expression of GAS5 in AFs isolated from sham and IVDD rats (n = 3). QRT-PCR detected (E) The expression levels of GAS5 and miR-221-3p in Afs after transfection with GAS5 overexpression plasmid and an empty vector. (n = 3) (F) The expression levels of GAS5 and miR-221-3p in Afs were detected by qRT-PCR after transfection with si-GAS5 and si-NC (n = 3). (G) The potential binding sites between GAS5 and miR-221-3p predicted by ENCORI (n = 3). (H) Luciferase activity and (I) AGO2-RIP were employed to evaluate the binding between lncRNA GAS5 and miR-221-3p (n = 3). (^*p < 0.05, ^**p < 0.01, and ^***p < 0.001).