Hplc grade organic solvents

In this study, the mycoparasitic biocontrol Sphaerodes mycoparasitia SMCD 2220-01 strain deposited in IDAC under accession number 301008-01 (Public Health Agency of Canada—International Depositary Authority of Canada Collection, Winnipeg, Canada) has been used for decomposition and detoxification of Fusarium mycotoxins. Zearalenone (ZEN), deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), and 15-acetyl-deoxynivalenol (15-ADON) shown in Fig. 1 were purchased from Sigma-Aldrich Canada Ltd., Oakville, ON. HPLC grade organic solvents were purchased from Fisher Scientific. The stock solutions of each of mycotoxins were prepared by dissolving each mycotoxin in acetonitrile. Potato dextrose broth (PDB, BD Difco) and agar (PDA) were used for maintaining SMCD 2220-01 and biodegradation experiments.Fig. 1

Chemical structure of the mycotoxin Zearalenone (ZEN), deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), and 15-acetyl-deoxynivalenol (15-ADON)

HPLC-grade organic solvents (Fisher Scientific, Ottawa, ON, Canada) and distilled deionized water were used throughout the experiment. Bacterial lipids were extracted following the Bligh-and-Dyer method^{15 (link)} with glass syringes, glass tubes, and ice-chilled solvents throughout the extraction process as previously described^{37 (link)}. After adding chloroform for monophasic extraction and eventual phase separation enhanced by centrifugation at 1,300 rpm for 5 min, the initial chloroform-rich phase was collected and supplemented with 0.5 mL 0.5 M sodium chloride (NaCl). After shaking by hand for 1 min, phase separation was again enhanced by centrifugation at 1,300 rpm for 5 min. The final chloroform-rich phase, approximately 4.2 mL, was collected for storage at −80 °C.

Methoxy poly(ethylene glycol)-b-poly(lactide-co-glycolide) (mPEG-PLGA) (MW: 2,000–15,000 Da) and Poly(lactide-co-glycolide) (PLGA) (MW: 10,000–15,000 Da) were purchased from PolySciTech^® (West Lafayette, IN, USA). GSK461364A was obtained from APExBIO (Houston, TX, USA) and reconstituted in ethanol; BPD-MA (Verteporfin) was purchased from Sigma Aldrich (St. Louis, MO, USA); MTS assay kit was purchased from Promega Corporation^® (Madison, WI, USA), LIVE/DEAD^® Cell Imaging Kit was bought from Invitrogen™ (Carlsbad, CA, USA). Triton-X and other HPLC grade organic solvents were purchased from Fisher Scientific™ (Agawam, MA, USA). Annexin-V, Alexa Fluor™ 647 conjugate was purchased from Invitrogen™. Propidium iodide (PI), Annexin binding buffer and phosphate buffer saline (PBS) were purchased from abcam (Cambridge, MA, USA).
Human GBM cell line, U87-MG was purchased from ATCC^® (Manassas, VA, USA) and cultured in MEM alpha modification media with l-glutamine (Genclone™) with 10% fetal bovine serum (FBS) (Genclone™, San Diego, CA, USA), 1% PenStrep (Gibco™, Fisher Scientific) maintained at 37 °C, 5% CO₂.