Cell viability was evaluated by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay and by crystal violet staining as previously described (76 (link), 77 (link)). Briefly, the MTT assay was conducted using the CellTiter 96 nonradioactive cell proliferation assay (MTT) (Promega; Madison, WI, USA). Cells were plated in a 96‐well plate at a concentration of 3 × 106 cells/mL in 100 μL of medium. The MTT solution (15 μL) was added to each well, and after 2 to 4 h, the reaction was stopped by the addition of 100 μL of 10% SDS. Absorbance values were acquired using an Infinite 200 PRO (Tecan, Männedorf, Switzerland) multimode plate reader at a 570-nm wavelength.
For crystal violet staining, cells were fixed in 6% formaldehyde and incubated with 0.1% crystal violet for 15 min. Unbound staining was then washed with H2O, and cells were imaged using a Nikon Eclipse Ts2-FL microscope.
For crystal violet staining, cells were fixed in 6% formaldehyde and incubated with 0.1% crystal violet for 15 min. Unbound staining was then washed with H2O, and cells were imaged using a Nikon Eclipse Ts2-FL microscope.
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