Fig. 7

Bioenergetic profile in cells with modulated IGF2BP2 expression. Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were measured using a Seahorse 96XF Analyzer in HCT116 IGF2BP2 wildtype (WT) and knockout (KO) cells (A, B, E, F) or in HCT116 cells under control conditions (NE) or overexpressing IGF2BP2 (OE) (C, D). A-E: A Seahorse XF Cell Mito Stress Test was performed. After measuring basal OCAR and ECAR oligomycin was injected to shut down OXPHOS-dependent ATP production followed by adding FCCP as an encoupler to obtain the maximal mitochondrial respiration capacity. Rotenone/antimycine A shut down mitochondrial oxygen consumption by inhibiting respiratory chain complex I and III. Statistical analysis was performed using Student´s t-test. Data are shown as means ± SEM, n = 3 (4–8). F The Seahorse XF Real-Time ATP Rate Assay was conducted using the same concentration of oligomycin and rotenone/antimycine A (see methods). Statistical analysis was performed using Student´s t-test. Data are shown as means ± SEM; n = 3–4 (4–8). G Gene expression of the respiratory chain complex genes was performed by real-time RT-PCR analysis in HCT116 WT and HCT116 IGF2BP2 KO cells. Gene expression data were normalized to the housekeeping gene 18S. Data are shown as means ± SEM relative to the WT control; n = 3 (triplicates). Statistical analysis was performed using Student´s t-test