U251N cells were seeded in 96-well black plates (Costar, Corning, NY, USA) at 5,000 cells per well in 0.1 ml media and cultured for 24 h. Cells were treated with sahaquine (0.001, 1, 5, 10, 20, 25, 30, 40, and 50 µM), TMZ (0.001, 1, 10, 50, 100, 200, 300, 400, and 500 µM, Sigma-Aldrich, St. Louis, MO, USA), quercetin (0.001, 1, 10, 25, 50, 75, 100, 200, and 300 µM, Sigma-Aldrich, St. Louis, MO, USA), or SAHA (0.001, 0.1, 1, 2, 5, 8, 10, 25, and 50 µM, Cayman Chemical, Ann Arbor, MI, USA) for 24 or 72 h. Combination treatments included increasing concentrations of sahaquine (0.001, 1, 3, 5, 7, 10, 20, and 50 µM) with quercetin (100 µM), increasing concentrations of quercetin (0.001, 1, 10, 25, 50, 100, and 200 µM) with sahaquine (10 µM), TMZ (30 µM) with sahaquine (10 µM), quercetin (140 µM) or SAHA (1 µM), and buthionine sulfoximine (100 µM, Sigma-Aldrich, St. Louis, MO, USA) with sahaquine (10 µM) for 24 h or 72 h. Following treatment, cells were fixed with 4% paraformaldehyde (w/v, 10 min, BDH, Toronto, ON, Canada). Nuclei were labeled with Hoechst 33342 (10 µM, 10 min, Thermo Fisher Scientific, Eugene, OR, USA). Cells were washed with phosphate-buffered saline and imaged using a fluorescence microscope (Leica DMI4000B, Toronto, ON, Canada).
Zhang I., Beus M., Stochaj U., Le P.U., Zorc B., Rajić Z., Petrecca K, & Maysinger D. (2018). Inhibition of glioblastoma cell proliferation, invasion, and mechanism of action of a novel hydroxamic acid hybrid molecule. Cell Death Discovery, 4, 41.
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