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. 2009 Nov 2;119(12):3626–3636. doi: 10.1172/JCI39374

Figure 3. Bmi-1 induces PI3K/Akt activity and downregulates E-cadherin.

Figure 3

(A) Cell extracts were analyzed by immunoblotting with antibodies against the indicated proteins. (B) Half-life analysis of the Snail protein. Cells were treated with 20 μM cycloheximide (CHX) for the indicated times and then analyzed by Western blotting. (C) The effect of Bmi-1 on the subcellular localization of Snail proteins (green) was examined under a fluorescent microscope. Scale bar: 80 μm. (D) E-cadherin mRNA expression was examined by real-time RT-PCR in triplicate. Error bars represent SEM (n = 3). Rel. exp., relative expression. (E) Luciferase activity under the control of the human E-cadherin promoter construct was measured in the indicated cells in triplicate. Error bars represent SEM (n = 3). **P < 0.01. (F) Repression of E-cadherin by Bmi-1 is dependent on Snail. (G) Binding of Bmi-1 to the E-cadherin promoter at different Snail expression levels was analyzed by ChIP. The results display the means ± SEM for 2 experiments performed in triplicate.