Multiskan ascent 96 384 plate reader

The measurement was performed as previously described (Gómez-Durán et al 2011 (link)). Three cycles of freeze-thaw were performed on each sample. Each assay was performed on a Multiskan Ascent 96/384 Plate Reader (ThermoFisher Scientific, UK). To establish a normal control range, we performed the measurement on 12 control liver tissues. The activities of citrate synthase (CS) and complex II were measured by spectrophotometry as previously described (Gómez-Durán et al 2011 (link)). Complex IV activity was analysed as reduction of cytochrome c at 550 nm as previously described (Gómez-Durán et al 2011 (link)). All measured parameters were expressed in specific activity per mg/protein in the sample. Total protein was quantified by method of Bradford assay.

The levels of IL-6 and TNF-α were measured in both plasma and colon extraction using ELISA assay performed according to the manufacturer’s instructions (Thermo Fisher Scientific, Villebon-sur-Yvette, France). The optical density was measured at a wavelength of 450 nm using a microtiter plate reader (Multiskan Ascent 96/384 plate reader, Thermo Fisher Scientific, Villebon-sur-Yvette, France). The final results were expressed as pg/mL and the limit of detection of IL-6 and TNF-α were 4–500 pg/mL and 8–1000 pg/mL, respectively.