Fig. 2

BRD4 degrader ARV-825 inhibited mammosphere formation. A Molecular structure of BRD4 degrader ARV-825. B Proliferation assay using ARV-825 on MDA-MB-231 cells. The cells were cultured with increasing concentration range of ARV-825 for 24 h. C Effect of ARV-825 on BRD4 degradation. The cells were cultured using the indicated concentrations of ARV-825 for 24 h. D Effect of BRD4 degrader ARV-825 on mammosphere formation. ARV-825-treated mammosphere formation was reduced, as shown in the photos and graphs. Experiment values are represented as the mean ± SD of triplicates. E CSC marker, CD44⁺/CD24⁻ expression on ARV-825-treated cells. The cells were treated with ARV825 for 24 h. CD44⁺/CD24 expression was evaluated using a flow cytometer. F CSC marker, ALDH expression on ARV-825-treated cells. The cells were treated with ARV825 for 24 h. ALDH expression was measured using the ALDEFLUOR assay kit and a flow cytometer. G CSC-related gene expression on ARV-825-treated cells. The cells were treated with 0.5 µM ARV-825 for 18 h. The mRNA levels of CD44, c-Myc, OCT4, and SOX2 were measured by reverse-transcription quantitative polymerase chain reaction. H Inhibitory effect of ARV-825 on mammosphere growth. ARV825-treated mammospheres were divided into single cells, and equal numbers of cells were cultured. The number of cells was analyzed daily for 3 days by counting. Experiment values are represented as the mean ± SD of triplicates. Compared with the control as determined by student’s t-test or one-way ANOVA with Dunnett’s multiple comparisons tests, * p < 0.05