Fig. 4

LBPs abrogate YAP1-overexpression-induced augmentation of mitochondrial function and upregulation of related factor expression in breast cancer cells. (A,B) MMP changes were assessed with the JC-1 fluorescent probe in MDA-MB-231 cells after LBPs treatment, transfection with the YAP1-overexpression plasmid, or YAP1-siRNA. Data are presented as the mean ± SD (n = 3). *** P < 0.001, ns: not significant. Scale bar, 50 μm. (C,D) Representative fluorescence images illustrating ATP probe expression after transfection of pCMV-Mito-AT1.03 into MDA-MB-231 cells that had been subjected to LBPs treatment or transfected with the YAP1-overexpression plasmid, or YAP1-siRNA. Data are presented as the mean ± SD (n = 3). *** P < 0.001, ns: not significant. Scale bar, 50 μm. (E,F) RT-qPCR analysis of the relative mRNA expression of YAP1 and TAZ in MDA-MB-231 cells following LBPs treatment or transfection with the YAP1-overexpression plasmid or YAP1-siRNA. Data are presented as the mean ± SD (n = 3), ***P < 0.001, **P < 0.01, *P < 0.05, ns: not significant. (G) Western blot analysis of YAP1, TAZ, ACSL4 and GPX4 protein expression in MDA-MB-231 cells following LBPs treatment or transfection with the YAP1-overexpression plasmid or YAP1-siRNA. (H–K) The gray blots were analyzed with ImageJ software (n = 3). ***P < 0.001, **P < 0.01, *P < 0.05, ns: not significant. (L,M) Detection of mitochondrial Fe²⁺ levels using Mito-FerroGreen probe. Data are presented as the mean ± SD (n = 3), ***P < 0.001, ns: not significant. Scale bar, 100 μm.