Figure 3. CD9 and CD82 inhibition of... | Rockefeller University Press

Figure 3.

$Figure 3. CD9 and CD82 inhibition of Wnt/β-catenin signaling is GSK-3β independent. (A) CD9 and CD82 but not CD63 inhibit TOPflash activity stimulated by mutant S33Y–β-catenin. 24 h after transfection with the indicated plasmids, cells were harvested for TOPflash luciferase assay. Data are presented as the fold change as compared with the control condition (cells transfected with pcDNA3.1 empty vector alone). (B) CD9 and CD82 expression reduced cytosolic and nuclear pools of the mutant S33Y–β-catenin protein. Cell fractions prepared from HEK 293T cells transfected with the indicated plasmids were immunoblotted using anti-Xpress (β-catenin) and anti–β-actin antibodies. (C) Effects of GSK-3β inhibitors on β-catenin levels. 12 h after transfection with the indicated plasmids, HEK 293T cells were treated with 50 mM NaCl, 50 mM LiCl, 20 µM SB216763, or DMSO (control) for 12 h. Lysates were blotted with anti–β-catenin and anti-dephosphorylated β-catenin (anti-ABC). β-Actin was used as a loading control.$

CD9 and CD82 inhibition of Wnt/β-catenin signaling is GSK-3β independent. (A) CD9 and CD82 but not CD63 inhibit TOPflash activity stimulated by mutant S33Y–β-catenin. 24 h after transfection with the indicated plasmids, cells were harvested for TOPflash luciferase assay. Data are presented as the fold change as compared with the control condition (cells transfected with pcDNA3.1 empty vector alone). (B) CD9 and CD82 expression reduced cytosolic and nuclear pools of the mutant S33Y–β-catenin protein. Cell fractions prepared from HEK 293T cells transfected with the indicated plasmids were immunoblotted using anti-Xpress (β-catenin) and anti–β-actin antibodies. (C) Effects of GSK-3β inhibitors on β-catenin levels. 12 h after transfection with the indicated plasmids, HEK 293T cells were treated with 50 mM NaCl, 50 mM LiCl, 20 µM SB216763, or DMSO (control) for 12 h. Lysates were blotted with anti–β-catenin and anti-dephosphorylated β-catenin (anti-ABC). β-Actin was used as a loading control.

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