Fig. 5

CircECE1 regulates glucose metabolism via c-Myc. a-b. After knocking down c-Myc, over-expression of circECE1, the protein and RNA of genes related to glucose metabolism were assessed by real-time PCR (a) and WB (b). c. CircECE1 MUT decreased ATP production induced by CircECE1 WT in U2OS and 143B cells. d. Mitochondrial potential was decreased after CircECE1 MUT overexpression compared to that of CircECE1 WT. e. Glucose uptake and lactate production was lower in the CircECE1 MUT-overexpressing OS cells. f. CircECE1 MUT rescued the upregulated the glycolysis rate induced by CircECE1 WT, and this was supported by the ECAR. CircECE1 MUT rescued the upregulated OCR induced by CircECE1 WT. g. CircECE1 WT function in tumor cell proliferation as detected by CCK-8 assay. h. CircECE1 WT function in tumor cell proliferation as detected by EdU assay. Nuclei were stained with DAPI. The combined reaction between EdU and DAPI indicated cell that were in S phase. i. CircECE1 overexpression promoted cell growth based on the results of a colony formation assay (details are shown in the insets). Error bars represent the mean ± SD of three independent experiments. * P < 0.05. j. Cell migration abilities of U2OS and 143B cells transfected with CircECE1 WT and MUT or vector were evaluated using transwell migration assays. Data represent the mean ± SD (n = 3). * P < 0.05. Scale bar = 50 μm. k. CircECE1 MUT overexpression in osteosarcoma cells increased cell anoikis (n = 3) compared to that observed in response to CircECE1 WT