Fig. 3

Effect of IL-1RA expression on mitochondrial metabolism and reactive oxygen species (ROS) formation in OSCC cells. A Multiple status of mitochondrial metabolism was measured by oxygen consumption rate (OCR) using Agilent Seahorse XFe24 Analyzer via sequential delivery of the indicated mitochondrial modulators (oligomycin, FCCP, and a mixture of rotenone and antimycin A) to OECM-1 cells with overexpression of IL-1RA and control cells. B–C Basal OCR indicated by the slope (∆RFU/∆min) was measured with CLARIOstar Plus multi-mode plate reader using a commercial oxygen consumption-labeling kit, for HSC-3 cells with knockdown of IL-1RA and control cells in (B), and OECM-1 cells with overexpression of IL-1RA and control cells in (C). D–E ROS formation was measured by flow cytometry after DCFDA labeling of OECM-1 cells with overexpression of IL-1RA and control cells in the presence or absence of rotenone in (D), and metformin or phenformin in (E). Data were presented as mean ± SD from three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001. RFU, relative fluorescence unit; shLuc, knockdown of firefly luciferase; shIL-1RA, knockdown of IL-1RA; EV, empty vector; IL-1RA-OE, overexpression of IL-1RA