Protocol to assess changes in mitochondrial respiration in living cultured cells infected with flaviviruses

Mitochondria are essential organelles involved in energy production, making them prime targets for flaviviruses, such as dengue and Zika viruses, to enhance viral replication. Here, we present a protocol to measure multiple respiratory parameters in living hepatocarcinoma cells infected with flaviviruses in combination with pharmacological treatments or genetic expression modulation. We detail steps for infecting cells with flaviviruses and measuring the oxygen consumption rates using Seahorse technology. For complete details on the use and execution of this protocol, please refer to Freppel et al.¹.