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. 2021 Nov 17;13(22):5757. doi: 10.3390/cancers13225757

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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PCAIs induce concentration-dependent cell death in A549 spheroids. (A) Cells were seeded in Nunclon Sphera 96-well, U-shaped-bottom microplates and allowed to grow and form compact spheroids overnight. They were then treated with the indicated concentrations of the respective PCAIs for 48 h. The spheroids were stained with AO/EO (5 μg/mL), and the fluorescent images were taken at 4× magnification using the Nikon Ti Eclipse microscope. (B) Graph showing concentration-dependent decreases in mean AO/EB intensity ratios (n = 3) indicating the viable to non-viable cells ratios in the spheroids. Statistical significance (* p < 0.05 and *** p < 0.001) was established using one-way ANOVA followed by Dunnett posttest comparisons.