Fig. 6

PGC-1α promotes autophagic and mitophagic activity in microglia. a Western blot analysis and quantification of the expression of the autophagy-related protein LC3 in LPS-primed BV2 cells after ATP stimulation for 3 h. b Assessment of autophagic flux in the presence or absence of the lysosomal inhibitor BAF (50 nM) and determination of LC3-II and SQSTM1 levels. c Representative images of mRFP-GFP-LC3 staining showing the autophagosome (yellow) and autolysosome (red-only) formation in the BV2 cells, with or without PGC-1α overexpression after OGD treatment. Bar: 10 μm. d Quantification of the number of yellow and red LC3 puncta. e Assessment of the autophagic flux rate. f Representative images of LC3-MitoTracker colocalization showing mitophagic events in the BV2 cells with or without PGC-1α overexpression after OGD treatment. Bar: 10 μm. g Quantification of the number of LC3-MitoTracker colocalized puncta. h Quantification of the percentage of LC3-MitoTracker colocalized puncta. i Quantification of the number of mitolysosomes. j Representative images of the formation of mitolysosomes in the BV2 cells with or without PGC-1α overexpression after OGD treatment. Bar: 20 μm. k Representative TEM images of mitolysosomes (red triangle) in the BV2 cells with or without PGC-1α overexpression after OGD treatment. Bar: 1 μm (upper panels), 0.5 μm (lower panels). l Quantification of the number of mitolysosomes. *p < 0.05, **p < 0.01; n = 6 per group