Fig. 5: Fen enhances autophagic flux and mitophagy in primary neurons.

a Representative images of primary neurons, with or without Fen treatment after 24 h, probed with Mitotracker (red) and Lysotracker (green). Arrows denote co-localization of mitochondria and lysosomes. b Western blotting of cultured neuronal lysates was performed using antibodies to LC3A/B (top panel) and quantitative analysis of the LC3A/B II/LC3A/B I ratio is shown (bottom panel); CCCP (10 µM) was used as an inducer of mitophagy. c Representative images of primary neurons, with or without Fen treatment after 24 h, stained with antibodies to PDH (green) and Ub (red). d Representative images of primary neurons, with or without Fen treatment over 24 h, stained with antibodies to PDH (green) and p62 (red). e Representative images of primary neurons, with or without Fen treatment over 24 h, stained with antibodies to LC3A/B (green), p62 (purple), and Mitotracker (red). f Western blotting of cultured neuronal lysates was performed using antibodies to PINK1 (left panel) and quantitative analysis of protein-band intensities is shown (right panel). g Representative images of primary neurons, with or without Fen treatment over 24 h, stained with antibodies to PDH (green) and PINK1 (red). For all quantitative/statistical analysis, *p < 0.05; **p < 0.01; and ***p < 0.001. All data are presented as means ± SEMs. Scale bars are 20 μm in a, c–e, and 3 μm in g.