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

Identification of the loop structure and expression levels of hsa

Figure 2. Identification of the loop structure and expression levels of hsa_circ_0007292. (A) Schematic showing that hsa_circ_0007292 is a circular structure formed from exons 3-8 of ATP5C1 mRNA; the backsplicing site of hsa_circ_0007292 was confirmed by Sanger sequencing. (B) Hsa_circ_0007292 could resist the digestion of RNase R in OPLL cells according to the results of qRT-PCR (n=3). (C) qRT-PCR experiment of the nuclear (Nuc) and cytoplasmic (Cyt) RNAs revealed that hsa_circ_0007292 was predominantly localized to the cytoplasm. U6 and β-actin were used as the internal control for nuclear RNA and cytoplasmic RNA, respectively (n=3). (D) FISH assays indicated that hsa_circ_0007292 was mainly localized to the cytoplasm. The probes of hsa_circ_0007292, U6, and 18S were marked with Cy3, and the nuclei were stained with DAPI. Scale bar, 100 mm. (E) Hsa_circ_0007292 was showed a higher expression in OPLL cells (n=8) than in non-OPLL cells detected by qRT-PCR(n=8). All experiments were performed at least three times. Data are expressed as the mean ± SD. ***p < 0.001.