Figure 6.

Decreased β-Catenin Level in Neuronopathic GD NPCs

(A) Representative immunofluorescence images of control, GD3, and GD2 NPCs stained with anti-β-catenin antibody (green) and with DAPI (blue). Cells were either left untreated or were treated with 0.24 U/mL rGCase for 5 days. Magnification 20×; scale bar, 100 μm.

(B) Representative immunofluorescence images of control, GD3, and GD2 NPCs stained with anti-β-catenin antibodies (green) and with DAPI (blue). Cells were either left untreated or were treated with proteasome inhibitor (PSI) for 18 hr. Magnification 40×; scale bar, 75 μm.

(C) Representative immunofluorescence images of control and GD2 NPCs stained with anti-active β-catenin antibodies (green) and with DAPI (blue). Magnification 20×; scale bar, 100 μm.

(D) Representative Western blot showing active β-catenin level in control and GD2 NPCs that were either left untreated or were treated with PSI for 18 hr as indicated. β-Actin was used as a loading control.

(E) Representative Western blot showing active β-catenin level in control and GD2 NPCs. Cells were either left untreated, treated with rGCase for 5 days, or treated with the Wnt activator CHIR for 3 days as indicated. β-Actin was used as a loading control. Bar graph below represents quantitation of active β-catenin level in NPCs with and without treatment. Data represent average ± SEM, n = 3–4 per group (compiled data are repeats from one GD2 patient). ^∗p < 0.05 between untreated GD2 and all groups as assessed by one-way ANOVA.