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. 2018 Jun 22;8:9575. doi: 10.1038/s41598-018-27810-6

Figure 5.

Figure 5

Estrogen enhances Twist expression via activation of AKT and NF-κB phosphorylation in ER+ breast cancer cells. (a) T47D and MCF-7 cells were pretreated with 0.5 μM Tamoxifen, 50 μM PI3K inhibitor LY294002, 10 μM IKK inhibitor Bay11–7082 or 10 μM MEK inhibitor PD 0325901 for 12 h, followed by stimulation of 10 nM E2 for 6 h or not. The protein levels of ERα, AKT, ERK1/2, NF-κB p65 and Twist as well as ERα, AKT, ERK1/2 and NF-κB p65 phosphorylation in different groups of T47D and MCF-7 cells with indicated treatments were measured by Western blot. (b) MDA-MB-231 cells were pretreated with 0.5 μM Tamoxifen and 50 μM LY294002 for 12 h, followed by stimulation of 10 nM E2 for 6 h or not. The protein levels of AKT and Twist as well as AKT phosphorylation on Ser473 were then detected by Western blot. (c) MCF-7 cells were pretreated with 20 μM RS102895 for 12 h, followed by stimulation of 20 ng/ml rhCCL2 for 24 h. The protein levels of MMP-9, VEGF, AKT and Twist as well as AKT phosphorylation were then detected by Western blot. Images are shown representatively from 3 independent experiments. Cropped images are from those samples and antibodies processed and run on different parts of gels. Full-length blots are displayed in Supplementary Fig. S4b.