96 well plate for suspension cells

Cells were plated at a density of 500 cells per well in a 96-well plate for suspension cells (Sarstedt, Nümbrecht, Germany) and treated for 14 days with G007-LK and/or TMZ. Control cells were treated with DMSO. Sphere formation was subsequently measured as the number of spheres in each well using an automated colony counter (Gelcount, Oxford Optronics, Abingdon, UK). Only spheres with a diameter > 50 µM were counted. Differences between the cells treated with G007-LK and DMSO were assessed with an unpaired two-tailed Student’s t-test (Excel 14.6.6, Microsoft Office), and differences between the cells treated with TMZ and those that underwent TMZ/G007-LK cotreatment were assessed by one-way ANOVA with Tukey’s multiple comparison test (Prism 5.0a, GraphPad Software).

Cells were plated at a density of 500 cells per well in a 96-well plate for suspension cells (Sarstedt, Nümbrecht, Germany) and treated for 14 days with G007-LK and/or TMZ. Control cells were treated with dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA). Proliferation was subsequently assessed using the Cell Proliferation Kit II XTT (Roche, Basel, Switzerland) according to the manufacturer’s protocol. The cells were incubated with the XTT solution overnight before the absorbance was analyzed using a microplate reader. Differences between the cells treated with G007-LK or TMZ and DMSO were assessed with an unpaired two-tailed Student’s t-test (Excel 14.6.6, Microsoft Office, Redmond, WA, USA), and differences between the cells treated with TMZ and those that underwent TMZ/G007-LK cotreatment were assessed by one-way ANOVA with Tukey’s multiple comparison test (Prism 5.0a, GraphPad Software, San Diego, CA, USA).