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. 2020 Feb 21;8:112. doi: 10.3389/fbioe.2020.00112

FIGURE 4.

FIGURE 4

NPs’ effect on inhibiting intracellular oligomerization of N-terminal acetylated and A53T mutant αSYN (A) Overall intracellular αSYN uptake with and without NP treatment was evaluated with Alexa488 labeled αSYN. Microglia were visualized with nuclei stain Hoechst 33342 (Blue) and microglial scavenger receptor CD36 (Red). Scale bar = 20 μm (B) BV2 microglia treated with monomeric αSYN (Ac- and A53T) and NPs, were lysed after 24 h incubation to detect intracellular αSYN content. Western blot was conducted with cell lysates, staining for αSYN and beta-actin. Oligomeric αSYN was detected inside monomeric αSYN-treated microglia. A significant reduction in oligomer content was observed in NP-treated microglia, indicating that [FAA:TA] NPs can lower intracellular αSYN aggregation. (C) Quantified αSYN fluorescence intensity representing overall αSYN uptake in microglia. Error bar shows standard error of n = 3; *P = 0.0216 by one-way ANOVA. (D,E) αSYN monomer, dimer and oligomer bands intensity was quantified and data shown in bar graph. In (D), for monomers: *P = 0.0156 for Ac-αSYN only vs. [FAA:TA] NP; for oligomers: *P = 0.04802 for Ac-αSYN only vs. [FAA:TA] NP and ***P = 0.0006 for Ac-αSYN only vs. Tannic acid by one-way ANOVA. In (E), for oligomers: *P = 0.0210 for A53T αSYN only vs. [FAA:TA] NP by one-way ANOVA.