Vps34 in1

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.

The following chemical inhibitors were used: Z-YVAD-FMK (10 μM; Calbiochem/Merck), wortmannin (1 μM; Sigma), GDC-0941 (5 μM; Calbiochem/Thermo), piceatannol (25 μM; Santa Cruz Biotechnology), cytochalasin D (5 μM; Sigma-Aldrich), and Vps34-IN1 (1 μM) and SAR405 (1 μM) (both from the Division of Signal Transduction Therapy Unit, University of Dundee, Dundee, Scotland).