Journal: Redox biology

Article Title: The E3 ubiquitin ligase TRIM31 is involved in cerebral ischemic injury by promoting degradation of TIGAR.

doi: 10.1016/j.redox.2021.102058

Figure Lengend Snippet: Fig. 4. TRIM31 deficiency maintained mitochondria homeostasis in brain of mice subjected to middle cerebral artery occlusion (MCAO). (A) TRIM31 deficiency improved ultrastructural defects of neuronal mitochondria from mice subjected to MCAO. (B–D) The effect of TRIM31 on the expression of protein related to mitochondrial dynamics, PGC-1α, MFN1, MEN2 and DRP1, in the mice brain after 24 h of MCAO. Scale bars: 10 μm. Data were presented as the mean ± SEM of 3 independent experiments. *p < 0.05 vs. indicated group.

Article Snippet: Primary antibodies against TRIM31 (12543-1-AP), G6PD (66 373-1-1 g), DRP1 (12957-1-AP), MFN1 (13798-1-AP), MFN2 (12186-1-AP) and PGC-1α (66 369-1-1 g) were from Proteintech (Wuhan, China).

Techniques: Expressing

Journal: International journal of molecular sciences

Article Title: 1,5-Anhydro-D-fructose Protects against Rotenone-Induced Neuronal Damage In Vitro through Mitochondrial Biogenesis.

doi: 10.3390/ijms22189941

Figure Lengend Snippet: Figure 5. Immunoblotting and quantitative immunoprecipitation evaluated the effects of 1,5-AF treatment on PGC-1α and AMPK proteins and activities in rotenone-treated PC12 cells. (a) Evaluation of PGC-1α protein level by immunoblotting. In rotenone-treated cells, PGC-1α protein expression was increased by 1,5-AF treatment. (b) Evaluation of PGC-1α acetylation assays. In rotenone-treated cells, deacetylated PGC-1α was increased by 1,5-AF treatment. (c) Total AMPK and phosphorylated AMPK were evaluated by immunoblotting. In rotenone-treated cells, the ratio of phosphorylated AMPK to total AMPK was increased by 1,5-AF treatment. All data are expressed as the mean ± standard error of the mean of quadruplicate experiments. * p < 0.05. 1,5-AF, 1,5-anhydro-D-fructose; ACC-Ly, acetylated lysine; DMSO, dimethyl sulfoxide; Met, metformin; pAMPK, phosphorylated AMP-activated protein kinase; PBS, phosphate-buffered saline; PGC- 1α, peroxisome proliferator-activated receptor-γ coactivator 1α; Rot, rotenone; tAMPK, total AMP-activated protein kinase; tPGC1a, total PGC-1α. Note: tPGC1a in panel (b) represents PGC-1α in samples immunoprecipitated with anti-PGC-1α antibody, whereas PGC1a in panel (a) represents PGC-1α in cell lysates that were not subjected to immunoprecipitation.

Article Snippet: In accordance with a published protocol [22], PGC-1α protein was immunoprecipitated with anti-PGC-1α antibody (Novus Biologicals) and agarose beads (Santa Cruz); PGC-1α levels and acetylation were then detected using specific antibodies for PGC-1α (Novus Biologicals) and ACC-Ly (Cell Signaling), respectively.

Techniques: Western Blot, Immunoprecipitation, Expressing, Saline

Journal: International journal of molecular sciences

Article Title: 1,5-Anhydro-D-fructose Protects against Rotenone-Induced Neuronal Damage In Vitro through Mitochondrial Biogenesis.

doi: 10.3390/ijms22189941

Figure Lengend Snippet: Figure 8. Effects of PGC-1α silencing on the mitochondrial protective activity of 1,5-AF in cultured PC12 cells. (a) Representative confocal images of MitoTracker staining in cells transfected with PPARGC1A small interfering RNA (siRNA; magnification: 100×, scale bar: 20 µm). (b) Transfec- tion with PPARGC1A siRNA inhibited the increase in MitoTracker intensity of 1,5-AF-treated cells. (c) Representative confocal images of MitoTracker staining in cells transfected with control siRNA (magnification: 100×; scale bar: 20 µm). (d) In cells transfected with control siRNA, treatment with 1,5-AF increased the MitoTracker intensity in DMSO-treated cells. All data are expressed as the mean ± standard error of the mean. * p < 0.05. 1,5-AF, 1,5-anhydro-D-fructose; DMSO, dimethyl sulfoxide; PBS, phosphate-buffered saline; siPGC1a, cells transfected with PPARGC1A siRNA.

Article Snippet: In accordance with a published protocol [22], PGC-1α protein was immunoprecipitated with anti-PGC-1α antibody (Novus Biologicals) and agarose beads (Santa Cruz); PGC-1α levels and acetylation were then detected using specific antibodies for PGC-1α (Novus Biologicals) and ACC-Ly (Cell Signaling), respectively.

Techniques: Activity Assay, Cell Culture, Staining, Transfection, Small Interfering RNA, Control, Saline

Journal: International Journal of Nanomedicine

Article Title: Polyoxometalates Ameliorate Metabolic Dysfunction-Associated Steatotic Liver Disease by Activating the AMPK Signaling Pathway

doi: 10.2147/IJN.S485084

Figure Lengend Snippet: POM inhibit cholesterol synthesis and activate the AMPK signaling pathway. ( a ) TC levels in liver samples. ( b ) Expression of liver precursor (p) or mature (m) SREBP2 was detected by Western blot. ( c ) Expression of liver SREBP2 and HMGCR by IHC. ( d ) Western blot analysis of total or phosphorylated AMPK (p-AMPK) in the liver and ( e ) HepG2 cells with POM treatment with PA/OA stimulation for 24 h. ( f – h ) HepG2 cells were pretreated with the AMPK inhibitor, Compound C (CC), or DMSO for 2 h, and then cells were exposed to PA/OA with or without POM for 24 h. ( f ) Western blot analysis of AMPK signaling. ( g ) TG contents in HepG2 cells. ( h ) Western blot examination of FASN and SCD1. *, P <0.05; **, P <0.01; ns, not significant versus corresponding control ( n = 3–5).

Article Snippet: Rabbit anti-SIRT1 (Cat# DF6033), HMGCR (Cat# DF6518), CPT-1α (Cat# DF12004), PGC-1α (Cat# AF5395) antibodies and phospho-NF-κB p65 (Cat# AF2006) phosphorylated antibody were obtained from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Expressing, Western Blot, Control

Journal: International Journal of Nanomedicine

Article Title: Polyoxometalates Ameliorate Metabolic Dysfunction-Associated Steatotic Liver Disease by Activating the AMPK Signaling Pathway

doi: 10.2147/IJN.S485084

Figure Lengend Snippet: POM inhibit inflammation and promote FFA β-oxidation in MCD-induced mice. ( a ) Staining of liver slices with Oil Red O. ( b ) Liver TG and ( c ) FFA levels. ( d ) Western blot detection of SIRT1, PPARα, PGC-1α, and total or phosphorylated AMPK (p-AMPK) in the liver. ( e ) mRNA expression levels of ACOX1 and CPT-1α determined by qRT–PCR. ( f ) Western blot detection of inflammation-related proteins in the liver. ( g ) qRT–PCR detection of mRNA expression of TNFα, IL-1β, and IL-6. ( h ) ELISA detection of serum TNFα, IL-1β, and IL-6 levels. *, P <0.05; **, P <0.01; versus corresponding control ( n = 5). ## , P <0.01; versus MCD group ( n = 5).

Article Snippet: Rabbit anti-SIRT1 (Cat# DF6033), HMGCR (Cat# DF6518), CPT-1α (Cat# DF12004), PGC-1α (Cat# AF5395) antibodies and phospho-NF-κB p65 (Cat# AF2006) phosphorylated antibody were obtained from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Staining, Western Blot, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Exercise improves muscle mitochondrial dysfunction-associated lipid profile under circadian rhythm disturbance

doi: 10.4196/kjpp.2024.28.6.515

Figure Lengend Snippet: Representative Western blot analyses reveal (A-D) decreased catalase and superoxide dismutase type 2 (SOD2) levels with no significant change in superoxide dismutase type 1 (SOD1) levels. (E-G) Increased apoptotic protein expression was observed. (H-K) Decreased expression of major proteins in the electron transport chain and (L-O) proteins governing mitochondrial biogenesis was presented. All data are presented as mean ± SEM. RCR, regular circadian rhythm; ICR, irregular circadian rhythm; Bcl2, B-cell lymphoma 2; Bax, Bcl2 associated X; NADH, reduced nicotinamide adenine dinucleotide; SDHB, succinate dehydrogenase subunit B; COX-I, cytochrome oxidase (COX) subunit I; AMPK, AMP-activated protein kinase; PGC-1α, peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1alpha; Tfam, mitochondrial transcription factor A. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 as determined using two-way ANOVA and Tukey’s post-hoc analyses (n = 6).

Article Snippet: The membranes were incubated with primary antibodies for two hours to overnight as follows: β-actin (Invitrogen, MA1-140); p-AMPK α1/2 (Millipore, #07-681); AMPK α1/2 (SCBT, sc-25792); peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1α; GenTex, GTX37356); mitochondrial transcription factor A (Tfam; SCBT, sc-166965); NADH:ubiquinone oxidoreductase subunit A9 (NADH; Abcam, ab110242); succinate dehydrogenase (SDH) subunits B (SDHB; ab14714, Abcam); cytochrome oxidase (COX) subunit I (COX-I; ab14705, Abcam); superoxide dismutase type 1 (SOD1; SCBT, sc-8637); superoxide dismutase type 2 (SOD2; SCBT, sc-18503); Catalase (SCBT, sc-271358); p-P53 (SCBT, sc-51690); P53 (SCBT, sc-55476); Bcl2 associated X (Bax; SCBT, sc-7480); B-cell lymphoma 2 (Bcl2; SCBT, sc-7382); brain and muscle ARNT-like 1 (BMAL1; SCBT, sc-365645); period circadian regulator 2 (PER2; Invitrogen, #PA5-100107).

Techniques: Western Blot, Expressing

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Exercise improves muscle mitochondrial dysfunction-associated lipid profile under circadian rhythm disturbance

doi: 10.4196/kjpp.2024.28.6.515

Figure Lengend Snippet: Western blot analyses reveal (A-D) increased antioxidant levels stimulated a (E-G) decrease in apoptotic protein expression. (H-K) Results exhibit heightened expression of proteins involved in the electron transport chain and (L-O) proteins governing mitochondrial biogenesis. All data are presented as mean ± SEM. EXT, endurance exercise training; SED, sedentary; RCR, regular circadian rhythm; ICR, irregular circadian rhythm; SOD1 & 2, superoxide dismutase type 1 & 2; Bcl2, B-cell lymphoma 2; Bax, Bcl2 associated X; NADH, reduced nicotinamide adenine dinucleotide; SDHB, succinate dehydrogenase subunit B; COX-I, cytochrome oxidase (COX) subunit I; AMPK, AMP-activated protein kinase; PGC-1α, peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1alpha; Tfam, mitochondrial transcription factor A. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 as determined using two-way ANOVA and Tukey’s post-hoc analyses (n = 6).

Article Snippet: The membranes were incubated with primary antibodies for two hours to overnight as follows: β-actin (Invitrogen, MA1-140); p-AMPK α1/2 (Millipore, #07-681); AMPK α1/2 (SCBT, sc-25792); peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1α; GenTex, GTX37356); mitochondrial transcription factor A (Tfam; SCBT, sc-166965); NADH:ubiquinone oxidoreductase subunit A9 (NADH; Abcam, ab110242); succinate dehydrogenase (SDH) subunits B (SDHB; ab14714, Abcam); cytochrome oxidase (COX) subunit I (COX-I; ab14705, Abcam); superoxide dismutase type 1 (SOD1; SCBT, sc-8637); superoxide dismutase type 2 (SOD2; SCBT, sc-18503); Catalase (SCBT, sc-271358); p-P53 (SCBT, sc-51690); P53 (SCBT, sc-55476); Bcl2 associated X (Bax; SCBT, sc-7480); B-cell lymphoma 2 (Bcl2; SCBT, sc-7382); brain and muscle ARNT-like 1 (BMAL1; SCBT, sc-365645); period circadian regulator 2 (PER2; Invitrogen, #PA5-100107).

Techniques: Western Blot, Expressing

Journal: Physiological Research

Article Title: Ellagic Acid Attenuates Muscle Atrophy in STZ-Induced Diabetic Mice

doi: 10.33549/physiolres.934918

Figure Lengend Snippet: Effects of EA on mitochondrial function in gastrocnemius. (A) NRF-1 and (B) PGC-1α levels were detected by Western blot. (C) NRF-1 and (D) PGC-1α relative expressions were quantized. ** p<0.01 vs. CON; ## p<0.01 vs. DM.

Article Snippet: The antibodies of Atrogin-1, GRP-78, MuRF-1, NRF-1 and PGC-1α were obtained from Sangon Biotech (Shanghai, China).

Techniques: Western Blot