Figure 5.
The Aptamers of F5R1 and F5R2 Enhanced Lysosomal Degradation of α-syn and Rescued the Cell Defects
(A) SK-N-SH cells pre-treated with F5R1, F5R2 or random DNA sequence were transfected the α-syn or vector control vectors and incubated for 24 hr. The extracts were separated by SDS-PAGE and blotted onto PVDF membrane. The membrane was blocked and probed with the α-syn specific polyclonal antibody. β-actin served as the loading control. (B) Quantitative analysis of the total protein level of α-syn from (A). (C) SK-N-SH cells were similarly treated as in (A) except for incubation time (48 hr). The cell extracts were immunoblotted with the α-syn polyclonal antibody. β-actin served as the loading control. (D) Quantitative analysis of the total protein level of α-syn from (C). Data are presented as the mean ± SD (one-way ANOVA) ***p < 0.001 compared with vector group; ##p < 0.01 compared with α-syn overexpression group (n = 3). (e) Cells were treated similarly as in (C), except for treated with NH4Cl or MG132 after 6 hr of transfection. The cell extracts were immunoblotted with the α-syn polyclonal antibody. (F). Quantitative analysis of the total protein level of α-syn from (E). Data are presented as the mean ± SD (one-way ANOVA) **p < 0.01 compared with α-syn+F5R1 group; ###p < 0.001 compared with α-syn+F5R2 group (n = 5). (G and H) Cells were treated similarly as in (E), Forty-eight hr after transfection, the cell viability was assessed by MTT assay (G) and LDH assay (H). Data are presented as the mean ± SD (one-way ANOVA) ***p < 0.001 compared with vector group; ###p < 0.001 compared with α-syn overexpression group (n = 6).