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Figure 6 | Molecular Neurodegeneration

Figure 6

From: Autophagy is involved in oligodendroglial precursor-mediated clearance of amyloid peptide

Figure 6

Autophagy was involved in the degradation of Aβ 42 . A. NG2 cell line was treated with Aβ42 for specified times. The expression of LC3-II, and beclin1 was determined by immunoblotting. B. NG2 cell line was incubated with HiLyte Fluor™ 488-labeled Aβ42 (400 nM) for 24 hours, and immunostained using an anti-LC3-II antibody. Cell nuclei were labeled with DAPI. Scale bars = 5 μm. C. NG2 cell line was transiently transfected with a mCherry–LC3 plasmid, and autophagosomes were demonstrated by mCherry–LC3 puncta. Scale bars = 5 μm. D . NG2 cell line was exposed to Aβ42 for 6 hours, and then treated with wortmannin at the indicated concentrations for 18 hours. Aβ42, and the expression of LC3, and beclin1 was determined by immunoblotting (top panel). The results were quantified (bottom panel). E . NG2 cell line was exposed to Aβ42 for 6 hours, then treated with bafilomycin A1 (0.2 nM) for 18 hours. Aβ42, the expression of LC3, and beclin1 was determined by immunoblotting (top panel). The results were quantified (bottom panel). F. Top panel: NG2 cell line treated with specific beclin1 siRNA oligonucleotides (siBeclin1-1, siBeclin1-2, siBeclin1-3), scramble RNA oligonucleotides for 24 hours. The expression of beclin1 was analyzed by immunoblotting. Ct: no treatment; Scr: scramble RNA oligonucleotides. Middle panel: NG2 cell line was transfected with beclin1 siRNA for 24 hours, then treated with Aβ42 for another 24 hours. Aβ42, the expression of LC3, and beclin1 was determined by immunoblotting. Bottom panel: Results were quantified. All data were presented as mean ± SD; *p<0.05, **p<0.01, denote significant difference. All the experiments were replicated three times.

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