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Figure 4 | Molecular Brain

Figure 4

From: Lysosomal enzyme cathepsin D protects against alpha-synuclein aggregation and toxicity

Figure 4

CD reduces α-syn aggregation. a. CD reduces α-syn aggregation in an aggregation assay. 50% of the cells transfected with α-syn-GFP, synphilin and empty vector exhibited visible α-syn aggregates. Co-transfection of CD together with α-syn-GFP and synphilin reduced the number of cells with visible α-syn aggregates to 20%. n = 3 independent transfection, each in quadruplicate. *p < 0.05 compared to absence of exogenous CD by Student t-test. b. Double immunostaining for CD and α-syn in the aggregation assay after transfection of H4 cells with α-syn+synphilin+vector and α-syn+synphilin+CD. Many more cells transfected with synphilin+α-syn+vector have α-syn aggregates than cells transfected with synphilin+α-syn+CD. α-syn = red; CD = green. Higher magnification images of the boxed areas are shown at the left-most panel. α-syn aggregates tended to be found in cellular areas with low levels of CD staining, in H4 cells with either α-syn+synphilin+vector or α-syn+synphilin+CD. Scale bar = 25 micron. Arrows point to α-syn aggregates. c. Double immunostaining for CD and the V5 tag of synphilin in the aggregation assay after transfection of H4 cells with α-syn+synphilin+vector and α-syn+synphilin+CD. Many more cells transfected with synphilin+α-syn+vector have α-syn aggregates than cells transfected with synphilin+α-syn+CD. Synphilin = red; CD = green. The levels of synphilin immunostaining were indistinguishable in cells transfected with synphilin+α-syn+vector versus cells transfected with synphilin+α-syn+CD. This is true in cells with or without aggregates. Scale bar = 25 micron. Arrows point to α-syn aggregates.

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