Fig. 2

Calycosin inhibits cell proliferation and survival by regulating ERα, RASD1, and miR-375 in ER+ breast cancer cells. MCF-7, T47D, SKBR3, MDA-MB-468, and MCF10A cells were treated for 48 h with calycosin (0, 16 μM), 16 μM calycosin and premiR-375, or 16 μM calycosin and miR-375 siRNA. The protein and mRNA expression levels of (a, b) ERα, (a, c) RASD1, and (d) miR-375 were determined using Western blotting and qRT-PCR. The ERα and RASD1 protein expression levels were normalized to β-actin expression, the internal control. Transcript levels were normalized to those of U6 snRNA in the case of miR-375. e Cell proliferation was analyzed using the CCK-8 assay. Representative data from three independent experiments are shown. *p < 0.05 vs. control (0 μM); #p < 0.05 vs. 16 μM calycosin alone