Research Paper Volume 13, Issue 16 pp 20192—20217

Hsa-circ-0007292 promotes the osteogenic differentiation of posterior longitudinal ligament cells via regulating SATB2 by sponging miR-508-3p

MiR-508-3p represses SATB2 expression by targeting the 3’-UTR of SATB2. (A) Schematic exhibiting the 28 overlapping target mRNA of miR-508-3p predicted by the miRTarbase, Targetscan, and StarBase databases. The 28 mRNAs further overlapped with those related to the Go terms GO:0001503 and GO:0001649. (B, C) The expression levels of SATB2 in OPLL cells and non-OPLL cells were detected by qRT-PCR (n=8) and Western Blot (performed 3 times in 3 pairs of the samples) assays. (D) Schematic of the predicted target site between miR-508-3p and the 3’-UTR of SATB2 from the Targetscan database. (E) The luciferase reporter system assays showed that compared with negative control, the miR-508-3p mimic obviously suppressed the luciferase activity of the WT-SATB2 luciferase reporter vector, while the luciferase activity of MUT-SATB2 was not affected by miR-508-3p mimic(n=3). (F, G) qRT-PCR assays indicated that the expression level of SATB2 was significantly enhanced or suppressed by the miR-508-3p inhibitor or mimic(n=3). (H, I) MiR-508-3p inhibitor or mimic could promote or suppress SATB2 protein expression, respectively(n=3). All tests were conducted at least three times. Data are expressed as the mean ± SD. *P

Figure 7. MiR-508-3p represses SATB2 expression by targeting the 3’-UTR of SATB2. (A) Schematic exhibiting the 28 overlapping target mRNA of miR-508-3p predicted by the miRTarbase, Targetscan, and StarBase databases. The 28 mRNAs further overlapped with those related to the Go terms GO:0001503 and GO:0001649. (B, C) The expression levels of SATB2 in OPLL cells and non-OPLL cells were detected by qRT-PCR (n=8) and Western Blot (performed 3 times in 3 pairs of the samples) assays. (D) Schematic of the predicted target site between miR-508-3p and the 3’-UTR of SATB2 from the Targetscan database. (E) The luciferase reporter system assays showed that compared with negative control, the miR-508-3p mimic obviously suppressed the luciferase activity of the WT-SATB2 luciferase reporter vector, while the luciferase activity of MUT-SATB2 was not affected by miR-508-3p mimic(n=3). (F, G) qRT-PCR assays indicated that the expression level of SATB2 was significantly enhanced or suppressed by the miR-508-3p inhibitor or mimic(n=3). (H, I) MiR-508-3p inhibitor or mimic could promote or suppress SATB2 protein expression, respectively(n=3). All tests were conducted at least three times. Data are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P<0.001.