FIG. 4.

Stable LMP2A expression results in increased cytosolic and nuclear accumulation of endogenous β-catenin. HFK cells stably expressing HA-tagged LMP2A or Babe vector were deprived of supplements (EGF and BPE) and incubated with 25 μM LY or DMSO control for 24 h prior to harvest and fractionation into cytosolic and nuclear components. Western blot analysis with an antibody that recognizes endogenous β-catenin was performed. Anti-HA, anti-GRP78, and anti-actin Western blots were included to show the corresponding LMP2A levels, attest to the purity of the fractionation procedure, and serve as a loading control, respectively. Densitometry with the Image J software was performed to normalize β-catenin levels to the corresponding actin levels. Fold increase (or decrease) of the normalized β-catenin values relative to the HFK Babe plus DMSO controls are indicated below the β-catenin immunoblot.