The Combination of β-Asarone and Icariin Inhibits Amyloid-β and Reverses Cognitive Deficits by Promoting Mitophagy in Models of Alzheimer’s Disease

Figure 5

Combination drug intact autophagy and mitophagy. (a–c) APP/PS1 mouse. (a) Representative set of electron microscopy images showing autophagic vacuoles-autophagosomes (AU) and multilamellar bodies (MLB) (arrows) (scale bar 1 μm and magnification 23,000x). (b) Representative Beclin-1, LC3, p62, PINK1, Parkin, and p-Parkin levels with WB and (c) showed quantification of protein expression levels. (d–i) PC12 cells. (d) Beclin-1, LC3, and p62 levels with WB and (e) quantification of protein expression levels. (f) IF assay of Beclin-1, LC3, and p62 was detected after the treatment (scale bar 100 μm and magnification 400x). SABC-DyLight marker protein expression, DAPI marker cell nucleus, also with a merge paper and (g) showed quantification of protein expression levels. (h, autophagy) Fluorescence images of autophagic cells. DALGreen was utilized for the imaging of autophagic cells. Starved cells were observed in the culture medium and (i) the data quantification (scale bar 100 μm and magnification 630x) (Ex: 488 nm, Em: 500-563 nm). (h, Mtphagy) Fluorescence images of mitophagy cells with mitophagy staining (scale bar 100 μm and magnification 630x) (Ex: 561 nm, Em: 650 nm). Lysosomes, Ex: 488 nm, Em: 502-554 nm. Mitochondrial staining with Mitobright Deep Red, Ex: 640 nm, Em: 656-700 nm. (i) Quantification of the data. Compared with the control group, , , and ; compared with the AD group, ^#, ^##, and ^###; one-away ANOVA. At least three experiments were repeated independently with similar results. Full scans of all the blots are in the Supplementary Note.

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