Captisol

PLX3397 (Selleck, S7818, USA) were dissolved in 10% DMSO and 90% 100 mg/mL Captisol (Selleck, S4592, USA) at a concentration of 100 μM, and were delivered to the cerebrospinal fluid of mice vial intracisternal injection (10 μL per injection) under isoflurane inhalation (once per day for consecutive two days).

Bufalin (3beta, 5beta-3, 14-Dihydroxy-bufa-20, 22-dienolide) was purchased from Tauto Biotech Co., Ltd., Shanghai, China, with >98% purity as determined by HPLC analysis. MK2206 and Captisol were from Selleck Chemicals. The aforementioned drugs were prepared in Dimethyl sulfoxide (DMSO), aliquoted and stored at −20 °C.

All animal experiments were approved by the Institutional Animal Care and Use Committee (Columbia University, protocol AC-AABQ5551). 5 × 10⁶ H460 cells per mouse in 400 μL of 0.9% saline were injected subcutaneously in the right hind flank of 4–6 weeks old female NSG mice. Tumor volume was quantified using calipers to measure the length, width, and height of each tumor (V = ½ × L × W²). When tumors reached an average size of 400mm³, mice were randomized and assigned to the different treatment groups. S. typhimurium was prepared from overnight culture, controlled for OD 0.1 and concentration of 4.5 × 10⁷ 0r 4.5 × 10⁸ CFU/mL CFU/mL and intratumorally injected at a concentration of 5 × 10⁸ cells per mL in 1X PBS with total volume of 20–40 μL per tumor^{11 (link),12 (link)}. 0.5 mL of 10 μM AHL was injected subcutaneously the day after bacterial treatment to induce therapeutic expression. MK2206 was dissolved in 30% captisol (Selleck Chemicals cat.no. S4592) in 1X PBS and injected in 240 mg/kg concentration by oral gavage, twice a week. Animal experiments were carried out independently at Danino lab and at The Oncology Precision Therapeutics and Imaging Core (OPTIC) in Columbia University. We confirm that all methods were performed in accordance with the relevant guidelines and regulations.