Ulk1/2+/+ and Ulk1/2−/− MEFs stably expressing pGF1-GAS-LUC were plated in a 96-well plate (three or five replicates of 5000 cells per well) and, 24 hours later, were maintained in serum-free RPMI 1640 medium overnight. Serum-starved cells were left untreated (control) or were treated with mouse IFN-γ (2.5 × 103 IU/ml) for 6 hours and then lysed using 1× Reporter Lysis Buffer (Promega). Luciferase assay substrate buffer (Luciferase Assay System no. E4030, Promega) was used as per the manufacturer’s instructions, and luciferase activities were measured using a Cytation 3 cell imaging multimode microplate reader (BioTek).

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