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. 2012 Jul 3;287(34):28552–28563. doi: 10.1074/jbc.M112.387878

FIGURE 3.

FIGURE 3.

A, RNAi-mediated depletion of β-catenin leads to loss of K63-polyubiquitin-modified APC. HEK293 cells were transfected with the indicated siRNAs for 72 h, followed by treatment with 10 μm MG132 together with vehicle (−) or Wnt3a (200 ng/ml) for 4 h. Lysates were immunoprecipitated with K63 polyubiquitin linkage-specific antibodies, and analyzed by immunoblot as indicated. B, HEK293 cells were transfected with siRNA for 72 h as indicated, followed by treatment with 10 μm MG132 for 4 h. Lysates were immunoprecipitated with K63 polyubiquitin linkage-specific antibodies, and analyzed by immunoblot as indicated. C, RNAi-mediated depletion of β-catenin leads to loss of APC-Axin interaction. HEK293 cells were transfected with the indicated siRNAs for 72 h, followed by treatment with 10 μm MG132 for 4 h. Lysates were immunoprecipitated with anti-APC and analyzed by immunoblot as indicated.