For mitochondrial depolarization, LZRS and YFP-Parkin cells were treated for 48 h with 12.5 μM CCCP (Sigma-Aldrich), 2 d after irradiation or etoposide treatment.
Antioxidant treatment was performed daily in irradiated cells treated 8 h before irradiation with 100 nM MitoQ (MedKoo Bioscience) for 10 d. Unless stated otherwise, for JNK inhibition cells were treated daily for 8 h before the irradiation with 20 μM SP600125 (Selleck Chemicals) for 10 d. For MRE11 inhibition, cells were treated with 100 μM Mirin (Selleck Chemicals) immediately after the irradiation for 10 d. The following HDAC inhibitors were used: 100 nM Trichostatin A (TSA) (Sigma- Aldrich) and Vorinostat (SAHA) (Selleck Chemicals) at the indicated doses were added 8 h before the irradiation for 10 d, unless stated otherwise. Drugs were replaced daily. Recombinant human IL1α protein (R&D Systems) was added to irradiated cells at 20 ng/mL for 24 h in serum-free medium.
Antioxidant treatment was performed daily in irradiated cells treated 8 h before irradiation with 100 nM MitoQ (MedKoo Bioscience) for 10 d. Unless stated otherwise, for JNK inhibition cells were treated daily for 8 h before the irradiation with 20 μM SP600125 (Selleck Chemicals) for 10 d. For MRE11 inhibition, cells were treated with 100 μM Mirin (Selleck Chemicals) immediately after the irradiation for 10 d. The following HDAC inhibitors were used: 100 nM Trichostatin A (TSA) (Sigma- Aldrich) and Vorinostat (SAHA) (Selleck Chemicals) at the indicated doses were added 8 h before the irradiation for 10 d, unless stated otherwise. Drugs were replaced daily. Recombinant human IL1α protein (R&D Systems) was added to irradiated cells at 20 ng/mL for 24 h in serum-free medium.