Infinite f200 pro multiplate reader

Stable infected Mel.7, Mel.17 and Mel.15 cells (n.s., sh-catu2) were treated with different concentrations of cisplatin, dacarbazine, paclitaxel or staurosporin for 48h, cell survival normalized to untreated cells (pos. contr.) and analyzed by CellTiter-Blue cell viability assay substrate. After 2 hours of incubation the cells were analyzed with Infinite F200 PRO multiplate reader (TECAN).
Caspase-Glo 3/7, 8 and 9 Assay from Promega was used to determine cells in apoptosis. Cells were treated as in the cell viability assay and luciferase substrate was added after 24 h. Luciferase was detected with Infinite F200 PRO multiplate reader (TECAN) and normalized for GFP values (stable infected).
JC-1—Mitochondrial Membrane Potential Assay Kit (Abcam) was used to investigate the level of cells in apoptosis. Stable infected Mel.7 cells (n.s., sh-catu2) were treated with different concentrations of cisplatin for 6 h and stained with JC-1 solution for 10 min. Relative fluorescence level was detected at a wavelength of 535 nm with Infinite F200 PRO multiplate reader (TECAN) and normalized for GFP values (488 nm).

HT29 cells were plated at 12,000 cells per well into 96-well plates. Cells were allowed to adhere for 24 h and serum-starved for an additional 24 h. Cells were then treated for 48 h with ZnCl₂ and/or ATRA (Stem Cell Technologies, Vancouver, BC, Canada), as indicated. Controls involved no ZnCl₂ or DMSO (0.1%; Fisher Scientific, Waltham, MA, USA) vehicle. After the indicated time, cells were washed in PBS and stained in 0.5% crystal violet (Sigma-Aldrich, St. Louis, MO, USA) solution for 20 min. Excess crystal violet was then washed off with deionized water and left to dry at room temperature for at least 24 h. Dried crystal violet stain was then re-suspended in 10% acetic acid solution and the optical density measured at 570 nm using a Tecan Infinite F200 Pro Multi-Plate Reader. Data were quantified using Tecan i-Control 1.8 software and analyzed using Microsoft Excel 365. Values reflect cell proliferation, as our standard curve showed that the absorbance values from crystal violet stain were directly proportional to cell number. The proliferation index was calculated by normalizing cell proliferation index of treated to untreated cells. Biological replicates were performed in triplicate with 6 technical replicates per condition, and the average was plotted with standard error of the mean (SEM) indicated.