Figure 3

Over-expression of α-synuclein attenuated the phagocytic ability of BV2 cells and lysosomal protein expression. BV2 cells were transiently transfected to express WT, A30P, or A53T α-synuclein for 48 hours. A) Transfected cells were incubated with FITC-labeled E. coli bioparticles (0.25 mg/mL) for 3 hours. After incubation, the media was removed and the signal from any unphagocytosed or membrane associated particles was quenched by incubating cells with a (0.25 mg/mL) trypan blue solution for 3 minutes. The fluorescence intensity of phagocytosed particles was measured via fluorescent plate reader (RFU). Each condition was performed with 8 replicates. B) Transfected cells were also lysed and Western blotted with anti-LAMP-1 and actin (loading control) antibodies. C) Optical density of LAMP-1 immunoreactive bands were normalized against their respective actin bands and averaged (± SD) from three independent experiments. * p < 0.05 compared to mock transfected cells, **p < 0.01 compared to mock transfected cells.