Additional file 2 of CircDOCK1 promotes the tumorigenesis and cisplatin resistance of osteogenic sarcoma via the miR-339-3p/IGF1R axis

Additional file 2: Figure S1. A: Volcano plot of genes in the GSE140256 dataset. B: Heatmap of the circRNA microarray (GSE140256) analysis results showing 10 dysregulated circRNAs, including 4 upregulated circRNAs, in OS tissues (log2FC > 1 or < − 1, P value < 0.05). C: CircRNA expression in OS cells. Figure S2. A: EdU incorporation assays and B: colony formation assays assessing the proliferation of U2OS cells transfected with circDOCK1-specific siRNAs or overexpression plasmids. C: Flow cytometric analysis assessing U2OS cell apoptosis. D-E: Transwell assay assessing the migration (D) and invasion (E) of U2OS cells. Figure S3. A: RIP assays detecting the binding between circDOCK1 and the 9 predicted RBPs. B: Heatmap of the RNA microarray (GSE36001) analysis results. C: Volcano plot of genes in the GSE36001 dataset. Figure S4. A-B: The transcript levels of circDOCK1 (A) and IGF1R (B) in xenograft tumors. C: The IGF1R and Ki-67 protein levels in xenograft tumors, as evaluated by IHC. D: H&E staining of mouse lungs after tail vein injection. Figure S5. A: Luciferase reporter assay detecting the binding of miR-339-3p to circDOCK1 and IGF1R in U2OS cell lines. B: CircDOCK1 and miR-339-3p transcript levels (left) as determined by RIP with an anti-AGO2 antibody in the U2OS cell line and AGO2 protein levels as determined by western blotting (right). C-D: Cotransfection of the miR-339-3p mimic and circDOCK1 overexpression plasmid or miR-339-3p inhibitor and circDOCK1 siRNA to measure the mRNA (C) and protein (D) levels of IGF1R in U2OS cell lines. E-I: Cotransfection of the miR-339-3p mimic and circDOCK1 overexpression plasmid to evaluate malignant transformation by CCK-8 (E), EdU incorporation (F), flow cytometry (G), Transwell migration (H) and Transwell invasion (I) assays in the U2OS cell line. Figure S6. A: Relative viability of circDOCK1 overexpression plasmid- or siRNA-transfected cells after 48 h of exposure to DOX at the specified concentrations. Figure S7. A-B: Cell growth (A) and apoptosis (B) in U2OS/DDP cell lines cotransfected with the miR-339-3p inhibitor and circDOCK1 siRNA. C: Tumor weights. D: IGF1R protein levels in xenograft tumors, as evaluated by IHC.