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Figure 4 | Molecular Cancer

Figure 4

From: Estrogen promotes stemness and invasiveness of ER-positive breast cancer cells through Gli1 activation

Figure 4

Knockdown of Gli1 in ER-positive breast cancer cells inhibited estrogen induced sphere formation and CD44+/CD24-/lowcells. (A) MCF-7 cells were transfected with control shRNA (shVEC) or shGli1 (shGli1-1 or shGli1-2) in the absence or presence of E2. Gli1 protein levels were analyzed using western blotting. β-Actin levels were measured as a loading control. (B) MCF-7 cells were treated with 10 nM E2 or ETOH as a control and transfected with shGli1-1, shGli1-2 or shVEC. After 4 days, cells were stained with anti-CD44-APC and anti-CD24-PE antibodies, and CD44+/CD24-/low subpopulations were examined using flow cytometry. (C) Histograms illustrate percentages of CD44+/CD24-/low subpopulations. (D) Representative images of MCF-7 and HCC1428 cells in the absence or presence of E2. MCF-7 and HCC1428 cells (1 cell/μL) were cultured in 96-well plates containing 100 μL SFM in each well with or without E2 for seven days. (E & F) The number of spheres was counted under a microscope. Error bars represent SEMs. **, ## indicate significant differences from the control (p < 0.001).

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