Cyclophosphamide

Five days postinjection, mice were primed with 10 mg·kg⁻¹ Gedatolisib or vehicle and then treated weekly (days 7, 14, 21, and 28 postinjection) with inhibitor +/− doxorubicin/Adriamycin and cyclophosphamide (AC). Gedatolisib stocks were made up fresh prior to each injection and injected via the lateral tail vein. Note that the drug must be injected slowly to avoid toxicity. Injection of 100 μL over approximately 5 s or more was appropriate to prevent injection toxicity. Vehicle was stored at 4 °C and injected in the same manner. One hour following inhibitor or vehicle treatment, AC [2 mg·kg⁻¹ Adriamycin/doxorubicin (Selleckchem S1208, in ddH20) and 60 mg·kg⁻¹ cyclophosphamide (Selleckchem S2057)] or vehicle control (5% DMSO, 30% PEG300, 5% Tween in ddH20) were delivered intraperitoneally. Bioluminescent imaging (BLI) was performed weekly on an IVIS Spectrum (Perkin Elmer) following intraperitoneal delivery of 100 μL d‐Luciferin (10 mg·mL⁻¹, BioVision, Inc.). The focal height was set to 0.7 cm.
To quantify DTCs remaining after treatment, one cohort of mice was euthanized on Day 29. A second cohort of mice was used to measure MFS over a prescribed course of 15 weeks.

Cytarabine, fludarabine, gemcitabine, cladribine, cyclophosphamide, doxorubicin and cisplatin were from Clinical Dept. of Hematology, University Hospital in Prague, Czech Republic. Temsirolimus, bortezomib, bendamustine and ibrutinib were purchased from Selleck Chemicals. Rituximab was kindly provided by Roche, Czech Republic.