Multiplex cytokine assay

Rat blood plasma was collected after 3 months of treatment and after addition of 1:100 diluted proteinase inhibitor cocktail (Millipore #539,134), samples were frozen at − 80 °C until testing. Multiplex cytokines in rat plasma were measured in duplicate using a Luminex platform (MAGPIX, Millipore, St. Charles, MO, USA) with the customized panel, MILLIPLEX MAP rat Cytokine/Chemokine Magnetic Bead (MILLIPLEX MAP kits, EMD Millipore, Billerica, MA, USA) and analyzed with MILLIPLEX analyst software (ViageneTech, Carlisle, MA, USA) according to the manufacturer's instructions. Plasma levels of granulocyte colony-stimulating factor (G-CSF), interleukin (IL)-1alpha (IL-1α), IL-4, IL-1beta (IL-β), IL10, IL-12(p70), interferon gamma (IFNγ) and IL17A were selected on the basis of significant upregulation resulting from mitochondrial transplantation⁷.