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

Journal: International Journal of General Medicine

Article Title: A Prospective Study of the Association of IL6 with the Critical Unit and Their Effect on in-Hospital Mortality in Critically Ill Patients

doi: 10.2147/IJGM.S474250

Figure Lengend Snippet: IL6 and the Critical Unit Markers of the Study Population

Article Snippet: Other serum proteins, such as Fission 1, Parkin, and S1P, were determined using commercially available enzyme-linked immunosorbent assay (ELISA) kits following manufacturers’ instructions (Fission 1: Abbexa, abx151559, Cambridge, UK; Parkin: Abcam, ab212159, Shanghai, China; S1P: MSKbio, kt99298, Wuhan, China).

Techniques:

Journal: International Journal of General Medicine

Article Title: A Prospective Study of the Association of IL6 with the Critical Unit and Their Effect on in-Hospital Mortality in Critically Ill Patients

doi: 10.2147/IJGM.S474250

Figure Lengend Snippet: Regression Models Between IL6 and Critical Unit Biomarkers of the Study Population

Article Snippet: Other serum proteins, such as Fission 1, Parkin, and S1P, were determined using commercially available enzyme-linked immunosorbent assay (ELISA) kits following manufacturers’ instructions (Fission 1: Abbexa, abx151559, Cambridge, UK; Parkin: Abcam, ab212159, Shanghai, China; S1P: MSKbio, kt99298, Wuhan, China).

Techniques:

Journal: The Journal of Headache and Pain

Article Title: SS-31 alleviated nociceptive responses and restored mitochondrial function in a headache mouse model via Sirt3/Pgc-1α positive feedback loop

doi: 10.1186/s10194-023-01600-6

Figure Lengend Snippet: Mitochondria were injured and mitochondrial homeostasis was altered after repeated IS infusion in a headache male mouse model. Male C57BL/6 mice were sham treated or dural-infused of inflammatory soup (IS) for 7 consecutive days, and then sacrificed for mitochondrial assessment. A - C Representative immunoblots and quantification of the protein levels of Pgc-1α ( **** p < 0.0001), Tfam ( *** p = 0.0009), Drp1 ( ** p = 0.0050), Mfn2, Fis1 ( * p = 0.0108), Beclin1, P62 ( ** p = 0.0021), Pink1 ( **** p < 0.0001), and Parkin in the TNC. n = 6 per group; Student’s t -test. D The levels of ATP ( ** p = 0.0022), MMP ( ** p = 0.0043), ROS ( *** p = 0.0002) and MDA ( * p = 0.0136) were detected and normalized by total protein concentrations in the TNC. n = 6 per group; Student’s t -test. E Mitochondrial ultrastructure in the TNC by TEM analysis. The abnormal mitochondria count percent ( *** p = 0.0002) and area percent ( **** p < 0.0001) were calculated. Red arrowhead, damaged mitochondria (swollen mitochondria and reduction of mitochondrial cristae). Black arrowhead, normal mitochondria. Scale bar, 1 μm. n = 4 per group; Student’s t -test. F Immunofluorescence staining of VDAC1 and nucleus (DAPI) in the TNC. The mean mitochondria length ( *** p = 0.0006) and area ( * p = 0.0285) were calculated. Scale bar, 5 μm. n = 6 per group; Student’s t -test. Data are represented as Mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 as compared to sham group

Article Snippet: The target protein information of the specific primary antibody was used as follows: Pgc-1α (1:1000, Novus Biologicals), mitochondrial transcription factor A (Tfam, 1:1000, Abcam), mitofusin 2 (Mfn2, 1:5000, Proteintech), fission 1 (Fis1, 1:2000, Wuhan Fine bio), dynamin related protein 1 (Drp1, 1:2000, Wuhan Fine bio), Beclin1 (1:1000, Wanleibio), PTEN-induced putative kinase 1 (Pink1, 1:1000, Wanleibio), Parkin (1:1000, Wanleibio), P62 (1:1000, Cell Signaling Technology), c-fos (1:5000, Proteintech), Sirt3 (1:1000, Novus Biologicals/ Wanleibio), proliferating cell nuclear antigen (Pcna, 1:500, Wanleibio), cytochrome oxidase subunit IV (CoxIV, 1:5000, Proteintech) and β-actin (1:3000, Servicebio).

Techniques: Western Blot, Immunofluorescence, Staining

Journal: The Journal of Headache and Pain

Article Title: SS-31 alleviated nociceptive responses and restored mitochondrial function in a headache mouse model via Sirt3/Pgc-1α positive feedback loop

doi: 10.1186/s10194-023-01600-6

Figure Lengend Snippet: SS-31 restored mitochondrial function and mitochondrial homeostasis in an IS-induced headache mouse model. C57BL/6 mice received sham, IS or SS-31 treatment for 7 consecutive days, and were sacrificed to evaluate the effects of SS-31 on mitochondrial function and mitochondrial homeostasis in TNC. A - C Western blot analysis was used to asses expression levels of Pgc-1α (F(2, 15) = 5.434, * p = 0.0168; * p = 0.0384, + p = 0.0249), Tfam (F(2, 15) = 6.457, ** p = 0.0095; * p = 0.0443, ++ p = 0.0100), Mfn2, Drp1, Fis1 (F(2, 15) = 15.74, *** p = 0.0002; ** p = 0.0015, +++ p = 0.0003), P62 and Pink1 (F(2, 15) = 5.143, * p = 0.0199; * p = 0.0153) in different groups. n = 6 per group; One-way ANOVA. D Western blot analysis and quantification of Sirt3 (F(2, 15) = 10.66, ** p = 0.0013; ** p = 0.0011, + p = 0.0208) protein level normalized to β-actin. n = 6 per group; One-way ANOVA. E The levels of ROS (F(2, 15) = 25.33, **** p < 0.0001; **** p < 0.0001, +++ p = 0.0003), ATP (F(2, 15) = 7.704, ** p = 0.0050; * p = 0.0413, ++ p = 0.0045), MMP (F(2, 15) = 25.89, **** p < 0.0001; **** p < 0.0001, +++ p = 0.0009) and MDA (F(2, 15) = 3.854, * p = 0.0446; + p = 0.0362) were detected and normalized by total protein concentrations in different groups. n = 6 per group; One-way ANOVA. (F) Mitochondrial ultrastructure in TNC by TEM analysis. The abnormal mitochondria count percent (F(2, 15) = 14.39, *** p = 0.0003; *** p = 0.0004, ++ p = 0.0034) and area percent (F(2, 15) = 13.70, *** p = 0.0004; *** p = 0.0003, + p = 0.0120) were calculated. Red arrowhead, damaged mitochondria (swollen mitochondria and reduction of mitochondrial cristae). Black arrowhead, normal mitochondria. Scale bar, 500 nm. n = 6 per group; One-way ANOVA. Data are represented as Mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 as compared to sham group. + p < 0.05, ++ p < 0.01 and +++ p < 0.001 as compared to IS group

Article Snippet: The target protein information of the specific primary antibody was used as follows: Pgc-1α (1:1000, Novus Biologicals), mitochondrial transcription factor A (Tfam, 1:1000, Abcam), mitofusin 2 (Mfn2, 1:5000, Proteintech), fission 1 (Fis1, 1:2000, Wuhan Fine bio), dynamin related protein 1 (Drp1, 1:2000, Wuhan Fine bio), Beclin1 (1:1000, Wanleibio), PTEN-induced putative kinase 1 (Pink1, 1:1000, Wanleibio), Parkin (1:1000, Wanleibio), P62 (1:1000, Cell Signaling Technology), c-fos (1:5000, Proteintech), Sirt3 (1:1000, Novus Biologicals/ Wanleibio), proliferating cell nuclear antigen (Pcna, 1:500, Wanleibio), cytochrome oxidase subunit IV (CoxIV, 1:5000, Proteintech) and β-actin (1:3000, Servicebio).

Techniques: Western Blot, Expressing

Journal: The Journal of Headache and Pain

Article Title: SS-31 alleviated nociceptive responses and restored mitochondrial function in a headache mouse model via Sirt3/Pgc-1α positive feedback loop

doi: 10.1186/s10194-023-01600-6

Figure Lengend Snippet: SS-31 restored mitochondrial function and mitochondrial homeostasis in H 2 O 2 -induced PC12 cells. A PC12 cells were treated with SS-31 (100 nM) for 2 h, followed by additional H 2 O 2 (300 nM) for 12 h. B Western blot analysis of Sirt3 and Pgc-1α (F(2, 6) = 7.438, * p = 0.0237; * p = 0.0452, + p = 0.0292). Protein levels of Sirt3 and Pgc-1α were quantified normalized to β-actin. n = 3 per group; One-way ANOVA. (C) Western blot analysis of Tfam (F(2, 6) = 6.981, * p = 0.0272; + p = 0.0254), Drp1, Fis1 (F(2, 6) = 8.292, * p = 0.0188; * p (H2O2) = 0.0200, * p (SS-31+H2O2) = 0.0485) and Pink1 (F(2, 6) = 20.93, ** p = 0.0020; ** p (H2O2) = 0.0037, ** p (SS-31+H2O2) = 0.0030). Protein levels were quantified normalized to β-actin. n = 3 per group; One-way ANOVA. D and E Flow cytometric analysis was used to examine MMP (F(2, 6) = 16.90, ** p = 0.0034; ++ p = 0.0045) and ROS (F(2, 6) = 7.564, * p = 0.0229; * p (H2O2) = 0.0314, * p (SS-31+H2O2) = 0.0388) levels in different groups. n = 3 per group; One-way ANOVA. Data are represented as Mean ± SD; * p < 0.05 and ** p < 0.01 as compared to con group. + p < 0.05 and ++ p < 0.01 as compared to H 2 O 2 group

Article Snippet: The target protein information of the specific primary antibody was used as follows: Pgc-1α (1:1000, Novus Biologicals), mitochondrial transcription factor A (Tfam, 1:1000, Abcam), mitofusin 2 (Mfn2, 1:5000, Proteintech), fission 1 (Fis1, 1:2000, Wuhan Fine bio), dynamin related protein 1 (Drp1, 1:2000, Wuhan Fine bio), Beclin1 (1:1000, Wanleibio), PTEN-induced putative kinase 1 (Pink1, 1:1000, Wanleibio), Parkin (1:1000, Wanleibio), P62 (1:1000, Cell Signaling Technology), c-fos (1:5000, Proteintech), Sirt3 (1:1000, Novus Biologicals/ Wanleibio), proliferating cell nuclear antigen (Pcna, 1:500, Wanleibio), cytochrome oxidase subunit IV (CoxIV, 1:5000, Proteintech) and β-actin (1:3000, Servicebio).

Techniques: Western Blot

Journal: The Journal of Headache and Pain

Article Title: SS-31 alleviated nociceptive responses and restored mitochondrial function in a headache mouse model via Sirt3/Pgc-1α positive feedback loop

doi: 10.1186/s10194-023-01600-6

Figure Lengend Snippet: The effects of SS-31 on mitochondrial function and homeostasis were partially counteracted by inhibiting Sirt3/Pgc-1α. C57BL/6 mice received PBS + sham + DMSO, PBS + IS + DMSO, SS-31 + IS + DMSO, SS-31 + IS + 3-TYP or SS-31 + IS + SR-18292 treatment for 7 consecutive days, followed by sacrifice to evaluate mitochondrial function and mitochondrial homeostasis in TNC. A ROS levels were examined in different groups and normalized by total protein concentrations. n = 6 per group; One-way ANOVA. F(4, 25) = 7.078, *** p = 0.0006; * p (PBS+IS+DMSO) = 0.0222, ** p (SS-31+IS+SR-18292) = 0.0029, + p (SS-31+IS+DMSO) = 0.0259, ## p (SS-31+IS+SR-18292) = 0.0034. B ATP levels were examined in different groups and normalized by total protein concentrations. n = 9 per group; One-way ANOVA. F(4, 40) = 4.753, ** p = 0.0031; ** p (PBS+IS+DMSO) = 0.0096, * p (SS-31+IS+3-TYP) = 0.0301, + p (SS-31+IS+DMSO) = 0.0393. C Western blot analysis showed that expression of Sirt3 (F(4, 40) = 13.71, **** p < 0.0001; **** p (PBS+IS+DMSO) < 0.0001, **** p (SS-31+IS+3-TYP) < 0.0001, **** p (SS-31+IS+SR-18292) < 0.0001, + p (SS-31+IS+DMSO) = 0.0153, ## p (SS-31+IS+3-TYP) = 0.0487, ## p (SS-31+IS+SR-18292) = 0.0216) and Pgc-1α (F(4, 40) = 15.15, **** p < 0.0001; *** p (PBS+IS+DMSO) = 0.0002, **** p (SS-31+IS+3-TYP) < 0.0001, **** p (SS-31+IS+SR-18292) < 0.0001, ## p (SS-31+IS+3-TYP) = 0.0044, ## p (SS-31+IS+SR-18292) = 0.0039) in different groups. n = 9 per group; One-way ANOVA. (D) Representative immunoblots and quantification illustrated the levels of Tfam (F(4, 40) = 9.868, **** p < 0.0001; *** p (PBS+IS+DMSO) = 0.0004, *** p (SS-31+IS+3-TYP) = 0.0001, *** p (SS-31+IS+SR-18292) = 0.0003, + p (SS-31+IS+DMSO) = 0.0417, # p (SS-31+IS+3-TYP) = 0.0177, # p (SS-31+IS+SR-18292) = 0.0363), Drp1 (F(4, 40) = 5.655, ** p = 0.0011; *** p (PBS+IS+DMSO) = 0.0004, * p (SS-31+IS+SR-18292) = 0.0492, + p (SS-31+IS+DMSO) = 0.0429), Fis1 and Pink1 in different groups. n = 9 per group; One-way ANOVA. E Immunofluorescence staining of VDAC1 and nucleus (DAPI) in TNC. The mean mitochondria length (F(4, 25) = 5.225, ** p = 0.0034; * p (PBS+IS+DMSO) = 0.0232, * p (SS-31+IS+3-TYP) = 0.0410, + p (SS-31+IS+DMSO) = 0.0312) and area (F(4, 25) = 6.964, *** p = 0.0007; * p (PBS+IS+DMSO) = 0.0396, ++ p (SS-31+IS+DMSO) = 0.0033, # p (SS-31+IS+3-TYP) = 0.0150, ## p (SS-31+IS+SR-18292) = 0.0047) were calculated. Scale bar, 5 μm. n = 6 per group; One-way ANOVA. Data are represented as Mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 as compared to PBS + Sham + DMSO group. + p < 0.05 and ++ p < 0.01 as compared to PBS + IS + DMSO group. # p < 0.05 and ## p < 0.01 as compared to SS-31 + IS + DMSO group

Article Snippet: The target protein information of the specific primary antibody was used as follows: Pgc-1α (1:1000, Novus Biologicals), mitochondrial transcription factor A (Tfam, 1:1000, Abcam), mitofusin 2 (Mfn2, 1:5000, Proteintech), fission 1 (Fis1, 1:2000, Wuhan Fine bio), dynamin related protein 1 (Drp1, 1:2000, Wuhan Fine bio), Beclin1 (1:1000, Wanleibio), PTEN-induced putative kinase 1 (Pink1, 1:1000, Wanleibio), Parkin (1:1000, Wanleibio), P62 (1:1000, Cell Signaling Technology), c-fos (1:5000, Proteintech), Sirt3 (1:1000, Novus Biologicals/ Wanleibio), proliferating cell nuclear antigen (Pcna, 1:500, Wanleibio), cytochrome oxidase subunit IV (CoxIV, 1:5000, Proteintech) and β-actin (1:3000, Servicebio).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining

Journal: Plant Physiology

Article Title: Peroxisome Function, Biogenesis, and Dynamics in Plants ^{1 [OPEN]}

doi: 10.1104/pp.17.01050

Figure Lengend Snippet: Peroxisome dynamics. Peroxisome biogenesis and division are coordinated by peroxins (numbered ovals) that coordinate peroxisomal membrane protein insertion into the ER or the peroxisomal membrane. After preperoxisomes bud from the ER, peroxisomes mature through import of matrix proteins. Peroxisomes can be degraded by pexophagy, a type of specialized autophagy. Dynamic peroxisome extensions (peroxules) assist peroxisome interactions with other organelles and can be associated with peroxisome division. PEX11 promotes peroxisome division together with a group of proteins (PMD1, FIS1, DRP) that also act in division of mitochondria or chloroplasts. PMP, peroxisomal membrane protein.

Article Snippet: The Arabidopsis paralogs of yeast FISSION1 ( Kemper et al., 2008 ), FIS1A and FIS1B, are tail-anchored membrane proteins acting in both mitochondrial and peroxisomal fission (for review, see Hu et al., 2012 ).

Techniques: Membrane