Mic 101 modular incubator

The LYCF cell line was kindly provided by Dr. You-Hua Huang (South China Agricultural University, Guangzhou). The LYCF cells were cultured in Leibovitz’s-15 Medium (Gibco, USA) supplemented with 10% (v/v) fetal bovine serum (Gibco, USA) and 200 µg/mL penicillin-streptomycin (Gibco) at 27 °C. For the hypoxic challenge, LYCF cells were cultured in a MIC-101 modular incubator (Billups Rothenberg Inc., USA) with 1% O₂ and 99% N₂ for 0, 3, 6, 12, 24, and 48 h.

The large yellow croaker fry (LYCF) cell line was kindly provided by Dr. You-Hua Huang, South China Agricultural University. The LYCF cells were cultured in Leibovitz’s-15 Medium (Gibco, USA) supplemented with 10% (v/v) fetal bovine serum (Gibco, USA) and 200 µg/mL of penicillin-streptomycin (Gibco, USA) at 27 °C. For the normoxic conditions, the cells were cultured in an incubator. For the hypoxic treatments, the cells were cultured under hypoxic conditions at 1% O₂ and 99% N₂ in a MIC-101 modular incubator (Billups Rothenberg, USA) for 0, 1, 3, 6, 12, 24, and 48 h. For the ROS modification experiments, the functions and concentrations of ROS scavengers (N-acetylcysteine (NAC) and elamipretide Szeto-Schiller-31 (SS-31)) and inhibitors (apocynin, diphenylene iodonium (DPI), and 5-hydroxydecanoate (5-HD)) are shown in Table 1. The following experimental groups were established: normoxia, hypoxia, hypoxia+NAC, hypoxia+DPI, hypoxia+apocynin, hypoxia+SS-31, hypoxia+5-HD, and hypoxia+apocynin+5-HD. All modulators were added to the medium 1 h prior to hypoxic stress.