Crystal violet solution aq

Cells were seeded in 96-well plates at 1×10⁴ cells/well in growth media and left overnight in the incubator for the cells to adhere. The following day cells were treated for 24 h with 0–100 µM OphA. After treatment, the media on the cells was removed and the cells were washed in PBS twice. Cells were fixed in 100 µl of 1% (v/v) glutaraldehyde (aq; Sigma-Aldrich) for 30 min and stained with 100 µl of 0.5% (w/v) crystal violet solution (aq; Sigma-Aldrich) for at least 1 h. The plate was washed with water, dried overnight and cells were solubilized using 150 µl of solubilising solution [1% (w/v) sodium dodecyl sulphate (SDS; Fisher Fisher Scientific) and 10% (v/v) acetic acid (Sigma-Aldrich)]. The absorbance of the solution was measured at 590 nm using a Tecan Infinite F200 microplate reader. Samples were blank corrected and expressed as a percentage of the control cell viability. Experiments were performed in triplicate and repeated on three separate occasions.

Cells were seeded in 96-well plates at 1 × 10⁴ cells/well in growth media and left overnight in the incubator for the cells to adhere. The following day cells were treated for 24 h with concentrations between 1 and 1000 μM citral. After treatment, the media on the cells was removed and the cells were washed in PBS twice. Cells were fixed in 100 μl of 1% (v/v) glutaraldehyde (aq; Sigma-Aldrich) for 30 min and stained with 100 μl of 0.5% (w/v) crystal violet solution (aq; Sigma-Aldrich) for at least 1 h. The plate was washed with water and dried overnight and cells were solubilized using 150 μl of solubilizing solution (1% [w/v] sodium dodecyl sulfate [Fisher Scientific]) and 10% (v/v) acetic acid (Sigma-Aldrich)]. The absorbance of the solution was measured at 590 nm using a Tecan Infinite f200 microplate reader. Samples were blank corrected and expressed as a percentage of the control cell viability. Experiments were performed in triplicates and repeated on three separate occasions.