Journal: Cells

Article Title: A Role for PGC-1a in the Control of Abnormal Mitochondrial Dynamics in Alzheimer’s Disease

doi: 10.3390/cells11182849

Figure Lengend Snippet: MutAPP triggers dynamic imbalance of mitochondrial fission and fusion. N 2 A cells were transfected with pCDNA or APPswe plasmid for 48 h. ( A ) EGFP-labeled APPswe was successfully overexpressed in N 2 A cells. Expression patterns and qualification of the fusion proteins, including ( B , D ) OPA1, ( F , I ) MFN1 and ( G , J ) MFN2, as well as the fission proteins, including ( L , N ) DRP1 and ( P , R ) FIS1 were studied with Western blot. Expression patterns and qualification of ( C , E ) OPA1, ( H , K ) MFN2, ( M , O ) DRP1 and ( Q , S ) FIS1 from the parietal cortex samples of 2×Tg-AD mice were also examined with immunohischemistry. Scale bars = 100 μm. ( T ) Representative electron microscope images showing stages of mitochondria in cortical neurons of WT and APP/PS1 animals. Scale bar = 0.5 μm. Yellow stars: the fissive stage of mitochondria. Significance levels were set at ** p < 0.01, *** p < 0.001 for noted differences between pCDNA and APPswe groups and WT and APP/PS1 groups. Tubulin was used as the loading control.

Article Snippet: Primary antibodies against EGFP (1:1000, Beyotime, cat # AG281, Shanghai, China), Flag (1:1000, abm, cat # G188, Zhenjiang, China), OPA1 (1:1000, Boster, cat # PB0773, Wuhan, China), MFN1 (1:1000, Boster, cat # PB0263, Wuhan, China), MFN2 (1:800, Bioss, cat # bs-23685R, Beijing, China), DRP1 (1:1000, Wanleibio, cat # WL03028, Shenyang, China), FIS1 (1:1000, Boster, cat # A01932-2, Wuhan, China), BAX (1:5000, Abcam, cat # ab32503, Cambridge, MA, USA), Bcl-2 (1:2000, Abcam, cat # ab182858, Cambridge, MA, USA), PGC-1a (1:1000, Bioss, cat # bs-1832R, Beijing, China), KIF5A Flag (diluted 1:1000, Bioword, cat # BS71526, Nanjing, China), KIF5B (diluted 1:1000, Wanleibio, cat # WL04906, Shenyang, China), and the horseradish peroxidase-linked antibodies (1:5000, Beyotime, goat anti mouse IgG, cat # A0216 and goat anti rabbit IgG, cat # A0208, Shanghai, China) were used to probe these blots.

Techniques: Transfection, Plasmid Preparation, Labeling, Expressing, Western Blot, Microscopy

Journal: Cells

Article Title: A Role for PGC-1a in the Control of Abnormal Mitochondrial Dynamics in Alzheimer’s Disease

doi: 10.3390/cells11182849

Figure Lengend Snippet: PGC-1α rescues mutAPP-triggered dynamic imbalance of mitochondrial fission and fusion. N 2 A cells were transfected with pEnCMV / Pgc-1alpha plasmid and plasmid-encoding APPswe for 48 h. ( A ) Flag-labeled PGC-1α and EGFP-labeled APPswe were successfully overexpressed in N 2 A cells. Expression patterns and qualification of the fusion proteins, including ( B , D ) OPA1, ( F , I ) MFN1 and ( G , J ) MFN2, as well as the fission proteins, including ( L , N ) DRP1 and ( P , R ) FIS1, were studied with Western blot. Expression patterns and qualification of ( C , E ) OPA1, ( H , K ) MFN2, ( M , O ) DRP1 and ( Q , S ) FIS1 from the parietal cortex samples of 2×Tg-AD mice treated with Vector/ Pgc-1alpha were also examined with immunohistochemistry. Scale bars = 100 μm. ( T ) Representative electron microscopes showing stage of mitochondria in cortical neurons of AAV-Vector- and AAV- Pgc-1alpha treated APP/PS1 mice. Scale bar = 0.5 μm. Yellow stars: the fissive stage of mitochondria. Significance levels were set at ** p < 0.01, *** p < 0.001 for noted differences between APPswe + pEnCMV and APPswe + Pgc-1alpha groups, or APP/PS1 + Vector and APP/PS1 + Pgc-1alpha groups. Tubulin was used as the loading control.

Article Snippet: Primary antibodies against EGFP (1:1000, Beyotime, cat # AG281, Shanghai, China), Flag (1:1000, abm, cat # G188, Zhenjiang, China), OPA1 (1:1000, Boster, cat # PB0773, Wuhan, China), MFN1 (1:1000, Boster, cat # PB0263, Wuhan, China), MFN2 (1:800, Bioss, cat # bs-23685R, Beijing, China), DRP1 (1:1000, Wanleibio, cat # WL03028, Shenyang, China), FIS1 (1:1000, Boster, cat # A01932-2, Wuhan, China), BAX (1:5000, Abcam, cat # ab32503, Cambridge, MA, USA), Bcl-2 (1:2000, Abcam, cat # ab182858, Cambridge, MA, USA), PGC-1a (1:1000, Bioss, cat # bs-1832R, Beijing, China), KIF5A Flag (diluted 1:1000, Bioword, cat # BS71526, Nanjing, China), KIF5B (diluted 1:1000, Wanleibio, cat # WL04906, Shenyang, China), and the horseradish peroxidase-linked antibodies (1:5000, Beyotime, goat anti mouse IgG, cat # A0216 and goat anti rabbit IgG, cat # A0208, Shanghai, China) were used to probe these blots.

Techniques: Transfection, Plasmid Preparation, Labeling, Expressing, Western Blot, Immunohistochemistry