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. 2013 Jun 7;8(6):e65556. doi: 10.1371/journal.pone.0065556

Figure 1. AIB1 regulates the morphologies of breast cancer cells.

Figure 1

(A) Light microscopic images showing the morphology of different breast cancer cells. (B) Expression levels of AIB1, ERα and E-cadherin in different breast cancer cells as detected by western blot analysis. Cell extracts were prepared from the different cell lines and probed with specific antibody against AIB1, ERα and E-cadherin or β-actin. (C) Light microscopic images showing EMT morphological changes induced in T47D cells after treatment with EGF (50 ng/ml) or E2 (20 nM) for 24 h. (D) Western blot analysis showing the levels of AIB1, ERα, E-cadherin and β-actin expressions in T47D cells after treatment with EGF or E2. (E) Coimmunoprecipitation showing AIB1 and ERα complex increased after treatment with EGF or E2. T47D cells treated without or with EGF (50 ng/ml) or E2 (20 nM) for 24 h were subjected to immunoprecipitation with anti-AIB1 or control IgG antibodies, followed by western blot analysis with anti-ERα and anti-AIB1 antibodies. (F) Light microscopic images showing T47D cells without or with AIB1 knockdown (shAIB1#1 and shAIB1#2), ERα knockdown (shERα) or both AIB1 and ERα knockdown. Cells were treated with the corresponding iRNA and then plated out in 50-mm dishes and incubated for 3 days before observing. (G) The cells from (F) were counted and plotted as percentage of clustered or scattered cells relative to total number of cells (400–500). (H) Western blot analysis showing the levels of AIB1, ERα, E-cadherin and β-actin proteins in T47D cells from (F).