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. 2024 Feb 20;12(3):470. doi: 10.3390/biomedicines12030470

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© 2024 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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NSL-YHJ-2-27 treatment disaggregates compact spheroids. MDA-MB-468 and MDA-MB-231 cells were seeded in 96U round bottom Nunclon Sphera Plate and allowed to grow for 48 h. Once compact spheroids were formed, they were treated with the indicated concentrations of NSL-YHJ-2-27 (0, 1, 2, 5, and 10 µM) and observed for 72 h. They were then stained with 10 µg/mL of AO/EB dye mixture at 72 h post-PCAIs-treatment and brightfield (BF) (A,B) and fluorescent (C,D) images were captured at 4× magnification using a Nikon Ti Eclipse microscope. Scale bar = 100 µm. The mean intensity ratios AO/EB equivalent to the viable-to-dead cells in the spheroids were determined (E,F) from three replicates. Statistical significance (*, p < 0.05; ***, p < 0.001) was determined by 1-way ANOVA with post hoc Dunnett’s test.

NSL-YHJ-2-27 treatment disaggregates compact spheroids. MDA-MB-468 and MDA-MB-231 cells were seeded in 96U round bottom Nunclon Sphera Plate and allowed to grow for 48 h. Once compact spheroids were formed, they were treated with the indicated concentrations of NSL-YHJ-2-27 (0, 1, 2, 5, and 10 µM) and observed for 72 h. They were then stained with 10 µg/mL of AO/EB dye mixture at 72 h post-PCAIs-treatment and brightfield (BF) (A,B) and fluorescent (C,D) images were captured at 4× magnification using a Nikon Ti Eclipse microscope. Scale bar = 100 µm. The mean intensity ratios AO/EB equivalent to the viable-to-dead cells in the spheroids were determined (E,F) from three replicates. Statistical significance (*, p < 0.05; ***, p < 0.001) was determined by 1-way ANOVA with post hoc Dunnett’s test.