Figure 1.

Knock-out (KO) of specific mitochondrial complex I β subcomplex subunit 10 results in reduced mitochondrial respiration; enhanced respiratory syncytial virus (RSV) virus production. Human embryonic kidney (HEK)293T cells and KO derivatives lacking Complex I β subcomplex subunit 4 (FB4^−/−) or 10 (FB10^−/−) [21] were (A,B) assessed for their mitochondrial bioenergetic activities using the Seahorse XF96 Extracellular Flux Analyzer. (A) An example of a typical oxygen consumption rate (OCR) obtained in these experiments. OCR was measured in real time upon sequential additions of ATP synthase inhibitor oligomycin (Oligo, 1 mM), proton ionophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP, 1 mM), mitochondrial complex III inhibitor antimycin A (Anti, 1 mM), and mitochondrial complex I inhibitor rotenone (Rot, 1 mM). (B) Mitochondrial respiratory parameters of basal and ATP-linked respiration were determined as previously [19,23]. (C) Cell lines were infected with RSV (multiplicity of infections (MOIs) indicated) for 24 h prior to analysis for cell-associated virus by plaque assays to determine infectious virus (plaque forming units pfu/mL). Results represent the mean ± SEM (n = 3 independent experiments, each performed in triplicate). ***p < 0.001 compared to the wild-type cells.