Figures & data
Figure 1 AC007207.2 is upregulated in OS tissue and cells.
Notes: (A) Identification of AC007207.2 through public data obtained from TARGET database; (B) qRT-PCR results of the top three high-expressed lncRNAs AC015726.2, AC078906.1 and AC007207.2, *P < 0.05, **P < 0.01, ***P < 0.001; (C) the expression level of AC007207.2 between normal and OS tissue, ***P < 0.001; (D) Kaplan–Meier survival curve indicated that high AC007207.2 patients have a worse prognosis; (E) FISH assay was performed to evaluate AC007207.2 expression in tumor and normal OS tissue.
Abbreviations: OS, Osteosarcoma; FISH, Fluorescence in situ hybridization.
Figure 2 AC007207.2 promote cell motility ability in OS cell lines.
Notes: (A) The knockdown efficiency of AC007207.2 was validated by qRT-PCR, **P < 0.01, ***P < 0.001; (B) transwell assay was performed in sh-NC and sh-AC007207.2 cells, ***P < 0.001; (C) scratch-healing assay was performed in sh-NC and sh-AC007207.2 cells.
Figure 3 AC007207.2 facilitate proliferation and inhibit apoptosis in OS cell lines.
Notes: (A) Colony formation assay was performed in sh-NC and sh-AC007207.2 cells, ***P< 0.001; (B and C) CCK8 assay was performed in sh-NC and sh-AC007207.2 cells, **P < 0.01; (D) EdU assay was performed in sh-NC and sh-AC007207.2 cells, ***P < 0.001; (E) flow cytometric analysis was performed in sh-NC and sh-AC007207.2 cells to evaluate the effect of AC007207.2 on cell cycle, **P < 0.01; (F) flow cytometric analysis was performed in sh-NC and sh-AC007207.2 cells to evaluate the effect of AC007207.2 on cell apoptosis, ***P < 0.001; (G) Western blotting was performed to evaluate the difference of proliferation-related molecular marker in sh-NC and sh-AC007207.2 cells.
Figure 4 AC007207.2 specifically bound to miR-1306-5p.
Notes: (A) FISH assay revealed that AC007207.2 is mainly localized in the cytoplasm; (B) the overexpression efficiency of miR-1306-5p mimics detected by qRT-PCR, **P < 0.01; (C and D) the relative expression level of AC007207.2 in NC mimics and miR-1306-5p mimics group, **P < 0.01; (E) the relative expression level of miR-1306-5p in NC and AC007207.2 knockdown group, **P < 0.01; (F) the predicted AC007207.2 binding sites in the region of miR-1306-5p and the corresponding mutant sequence were shown; (G and H) the relative luciferase activity in AC007207.2 wild and mutation type cells, **P < 0.01; (I) the correlation between AC007207.2 and miR-1306-5p detected by qRT-PCR; (J) RNA pulldown assay was performed with the biotin-AC007207.2 probe and anti-probe to enrich miR-1306-5p.
Figure 5 miR-1306-5p inhibit the proliferation and motility ability of OS cell lines.
Notes: (A and B) CCK8 assay performed in NC mimics and miR-1306-5p mimics cells, **P < 0.01; (C) Western blotting was performed to evaluate the difference of proliferation-related molecular marker in NC mimics and miR-1306-5p mimics cells; (D) transwell assay performed in NC mimics and miR-1306-5p mimics cells, ***P < 0.001.
Figure 6 SIRT7 is a target gene of miR-1306-5p.
Notes: (A) The correlation between SIRT7 and miR-1306-5p in OS tissue; (B) the correlation between SIRT7 and AC007207.2 in OS tissue; (C) the SIRT7 mRNA expression level in NC mimics and miR-1306-5p mimics cells, **P < 0.01; (D) the predicted SIRT7 binding sites in the region of miR-1306-5p and the corresponding mutant sequence were shown; (E and F) the relative luciferase activity in SIRT7 wild and mutation type cells, **P < 0.01; (G) SIRT7 protein level in NC mimics and miR-1306-5p mimics cells; (H) RNA pulldown assay was performed with the biotin-miR-1306-5p probe and anti-probe to enrich SIRT7; (I) RIP assay demonstrated that Ago2 protein binds more to SIRT7 and miR-1306-5p control lgG, **P < 0.01.
Figure 7 AC007207.2 enhances the progression of OS cells by miR-1306-5p/SIRT7 axis.
Notes: (A and B) The knockdown efficiency of miR-1306-5p and SIRT7 were detected by qRT-PCR, **P < 0.01; (C) colony formation assay performed in NC, miR-1306-5p inhibitor and SIRT7 rescue group, **P < 0.01; (D) CCK8 assay performed in NC, miR-1306-5p inhibitor and SIRT7 rescue group, **P < 0.01; (E) transwell assay performed in NC, miR-1306-5p inhibitor and SIRT7 rescue group, ***P < 0.001.
