Mrt68921

Crizotinib (PF-02341066, Selleckchem, S1068), Ceritinib (LDK378, Selleckchem, S7083) and Lorlatinib (PF-6463922, Selleckchem, S7536) were dissolved in DMSO at a stock concentration of 10 mM. Alectinib (CH5424802, Selleckchem, S2762) was dissolved in DMSO at a concentration of 2 mM. The above mentioned chemicals were stored at -80 °C. Bafilomycin A1 (Enzo-Life sciences, BML-CM110) was dissolved in DMSO at a stock concentration of 200 µM. Chloroquine (Sigma-Aldrich, C6628) was reconstituted in distilled water to 10 mM. Both inhibitors were stored at − 20 °C. For autophagic flux measurements, Bafilomycin A1 was used at a final concentration of 200 nM and added during the last 2–5 h of treatment depending on the method used. For the clonogenic assay, Chloroquine was added at a concentration of 15 µM and incubated for 48 hr prior to re-seeding. The ULK1 Inhibitor MRT68921 (Sigma-Aldrich, SML1644) was dissolved in water at a final concentration of 20 mM and stored at -80 °C. SAR-405 from ApexBio (A8883) and VPS34-IN1 (S7980) from Selleckchem were dissolved in DMSO to 50 mM and stored at -80 °C. For the LDH experiments, MRT68921 was used at a concentration of 7.5 µM and VPS34-IN1 at 5 µM. Water served as vehicle control for MRT68921, whereas DMSO (Sigma-Aldrich, D4540) was used as a vehicle control for all other drugs dissolved in DMSO.

HeLa cells were pre-treated with DMSO or 1 µM MRT68921 (SML1644, Merck) in culture medium, 1 h before the infection. Medium was replaced with serum-free media containing DMSO or 1 µM MRT68921 and cells were infected with Poliovirus (PV) at MOI 5 (MOI 1 and MOI 0,1 for qPCR) for 1 h. Inoculum was then removed, and cells were incubated in fresh media containing 2% FBS and 1 µM MRT68921 and/or 5 µM Vps34-IN1 or 50 µg/mL hydantoin (5-(3,4-dichlorophenyl)-5-methylimidazolidine-2,4-dione (EN300-21815)), or 2 mM Guanidine hydrochloride (Sigma) for 3 h to 8 h. Vps34-IN1(17392, Cayman chemicals) was added to the cell media at 1 h p.i. to not interfere with the endocytosis-based viral entry. MRT68921 was added at 1 h p.i. After addition, all inhibitors were kept in the cell media throughout the experiments.

SBI-0206965 (Sigma-Aldrich) and MRT-68921 (Sigma-Aldrich) were dissolved in DMSO and distilled water, respectively, to prepare the 10 mg/ml stock solution. AML-bearing mice were intraperitoneally injected with a dose of 20 mg/kg SBI-0206965 diluted in PBS containing 5% polyethylene glycol 400 (Sigma-Aldrich) and 5% Tween 80 (Sigma-Aldrich) or MRT-68921 diluted in PBS according to the schedule (Fig. 5a).