Rescue from a two hit, high-throughput model of neurodegeneration with N-acetyl cysteine

► The first dual hit model of synergistic proteotoxic stress was developed in N2a cells. ► Toxic, but not subtoxic MG132 exacerbated the response to a second MG132 hit. ► Toxic MG132 raised adaptive autophagic defenses. ► Two hits of toxic MG132 caused a synergistic loss of glutathione. ► N-acetyl cysteine protected against synergistic proteotoxic stress by three viability assays.