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. 2022 Feb 28;12(6):2598–2612. doi: 10.7150/thno.70581

Figure 7.

Figure 7

SMARCD3 knockdown blocks paclitaxel-induced FOXO3 binding on pluripotency factor genes and inhibits BCSC enrichment. A, MDA-MB-231 cells were transfected with vector encoding NTC or either of two shRNAs targeting SMARCD3 (#1 and #2), and immunoblot assay was performed. B-D, MDA-MB-231 NTC or SMARCD3 knockdown subclones were treated with vehicle (V) or 10 nM paclitaxel (P) for 72 h, and ALDH (B; mean ± SEM; n = 3), mammosphere (C; mean ± SEM; n = 4), and qPCR (D; mean ± SEM; n = 3) assays were performed; *p < 0.05, **p < 0.01, ***p < 0.001 vs. NTC-V; #p < 0.05, ##p < 0.01, ###p < 0.001 vs. NTC-P; ns, not significant. E-I, MDA-MB-231 NTC or SMARCD3 knockdown subclones were treated with V or 10 nM P for 72 h. ChIP was performed using antibodies against FOXO3 (E), H3K27me3 (F), H3K27ac (G), KDM6A (H), or p300 (I), followed by qPCR with primers flanking FOXO3 binding sites in the NANOG, SOX2 and KLF4 gene (mean ± SEM; n = 4); *p < 0.05, **p < 0.01, ***p < 0.001 vs. NTC-V; ##p < 0.01, ###p < 0.001 vs. NTC-P; ns, not significant.