Gdc 0994

Drugs were dissolved in DMSO and stored at −80, and aliquots were thawn prior to each use. Drugs were dissolved at concentrations resulting in in vitro experimental conditions 0.15% DMSO maximum in the culture medium. Drug stocks: 20 mg/mL regorafenib (R‐8024), 15 mg/mL erlotinib (E‐4007), 20 mg/mL vemurafenib (V‐2800), and 1 mg/mL BEZ‐325 (N‐4288) from LC laboratories (Woburn, MA, USA); 20 mg/mL selumetinib (HY‐50706), 10 mg/mL AZD‐4547 (HY‐13330, 10 mg/mL GDC‐0994 (HY‐15947), 20 mg/mL folinic acid (HY‐17556), and 5 mg/mL oxaliplatin (HY‐17371) from MedChemExpress (Monmouth Junction, NJ, USA); 10 mg/mL vatalanib (S1101) and 10 mg/mL crenolanib (S2730) from Selleck Chemicals (Houston, TX, USA); 10 mg/mL 5‐fluorouracil (F6627) from Sigma‐Aldrich; 4 mg/mL Zaltrap^® from Sanofi (Paris, France). Single drugs or premixed drug combinations were incubated with the 2D cell cultures for 24 hours or 72 hours, applied at day 1 postseeding and with the 3D cell cultures for 72 hours or 72 hours + 48 hours, applied at day 2 or day 2 + day 5 postseeding [19, 20]. The 48‐hour retreatment was performed by adding another volume of media containing a 1x concentration of drugs to not affect the final concentration in the wells.