Journal: Experimental Animals

Article Title: Suppression of USP2 in mouse skeletal muscle: a model of oxidative stress in muscle tissue

doi: 10.1538/expanim.25-0032

Figure Lengend Snippet: Chemical blockade of ubiquitin-specific protease 2 (USP2) enhances oxidative stress in C2C12 myotubes. Differentiated C2C12 cells were treated with 10 µ M ML364 or the vehicle (2 mM DMSO) for 12 h (A, B) or 6 h (C–F). (A) Lactate dehydrogenase (LDH) content of the culture supernatant and (B) protein level of cyclin D1. A representative image is shown (B, top). (C) Mitochondrial reactive oxygen species (ROS) levels, (D) the glutathione disulfide/glutathione (GSSG/GSH) ratio, and (E) protein level of uncoupling protein 2 (UCP2). A representative image is shown (E, top). (F) Activity of thioredoxin-disulfide reductase (TrxR), glutathione reductase (GR), glutathione peroxidase (GPx), and superoxide dismutase (SOD). The data presented are the mean ± SD of 6 wells. * P <0.05 vs. vehicle.

Article Snippet: Primary antibodies targeting cyclin D1 (#RM9104; Thermo Fisher Scientific, Waltham, MA, USA), lamin B (#SC6217; Santa Cruz Biotechnology, Dallas, TX, USA), UCP2 (#CSB-PA299698, Cusabio, Houston, TX, USA), and GAPDH (#016-25523, Fujifilm Wako) were used.

Techniques: Ubiquitin Proteomics, Activity Assay

Journal: Frontiers in Physiology

Article Title: Hypoxanthine Induces Muscular ATP Depletion and Fatigue via UCP2

doi: 10.3389/fphys.2021.647743

Figure Lengend Snippet: Primer sequences used for qRT-PCR.

Article Snippet: The following antibodies were used: rabbit anti-UCP2 antibody (1:1500, 162448, ZENBIO, China), rabbit anti-UCP3 antibody (1:1500, 162449, ZENBIO, China), mouse anti-MyHC I antibody (1:1000, BA-D5-s, DSHB, United States), mouse anti-MyHC IIb antibody (1:1000, BF-F3, DSHB, United States), and rabbit anti-β-actin antibody (1:5000; AP0060, Bioworld, China).

Techniques:

Journal: Frontiers in Physiology

Article Title: Hypoxanthine Induces Muscular ATP Depletion and Fatigue via UCP2

doi: 10.3389/fphys.2021.647743

Figure Lengend Snippet: Hx induces the activation of UCP2 in skeletal muscle. (A) mRNA expression of UCP2 and UCP3 in the GAS muscle after 4 weeks Hx treatment ( n = 8 mice per group). (B,C) Western blot (B) and quantification (C) of UCP2 and UCP3 content in the GAS muscle after 4 weeks Hx treatment ( n = 6 per group). Dividing lines indicate spliced bands from the same gel. (D) mRNA expression of UCP2 and UCP3 in the GAS muscle after acute Hx treatment ( n = 8 mice per group). (E,F) Western blot (E) and quantification (F) of UCP2 content in the GAS muscle after acute Hx treatment ( n = 6 per group). Dividing lines indicate spliced bands from the same gel. β-actin was used as loading control both in qRT-PCR and western blot. Values are presented as means ± SEM. ∗ P ≤ 0.05 according to the non-paired Student’s t -test between individual groups.

Article Snippet: The following antibodies were used: rabbit anti-UCP2 antibody (1:1500, 162448, ZENBIO, China), rabbit anti-UCP3 antibody (1:1500, 162449, ZENBIO, China), mouse anti-MyHC I antibody (1:1000, BA-D5-s, DSHB, United States), mouse anti-MyHC IIb antibody (1:1000, BF-F3, DSHB, United States), and rabbit anti-β-actin antibody (1:5000; AP0060, Bioworld, China).

Techniques: Activation Assay, Expressing, Western Blot, Control, Quantitative RT-PCR

Journal: Frontiers in Physiology

Article Title: Hypoxanthine Induces Muscular ATP Depletion and Fatigue via UCP2

doi: 10.3389/fphys.2021.647743

Figure Lengend Snippet: UCP2 is required for the Hx-induced muscular ATP deficiency and weakness. (A) Experimental scheme for the acute Hx treatment on m UCP2 KO mice. (B,C) Western blot (B) and quantification (C) of UCP2 content in the GAS muscle after acute Hx treatment ( n = 6 per group). Dividing lines indicate spliced bands from the same gel. (D) Weights lifting score after acute Hx treatment ( n = 8 mice per group). (E) Grip strength after acute Hx treatment ( n = 8 mice per group). (F–H) GAS muscle ATP (F) , glycogen (G) , and lactate (H) content of mice after acute Hx treatment ( n = 7–8 mice per group). (I) mRNA expression of genes related to the glycolysis of GAS muscle after acute Hx treatment ( n = 7–8 mice per group). β-actin was used as loading control both in qRT-PCR and western blot. Values are presented as means ± SEM; ∗ P ≤ 0.05 according to the non-paired Student’s t -test between individual groups.

Article Snippet: The following antibodies were used: rabbit anti-UCP2 antibody (1:1500, 162448, ZENBIO, China), rabbit anti-UCP3 antibody (1:1500, 162449, ZENBIO, China), mouse anti-MyHC I antibody (1:1000, BA-D5-s, DSHB, United States), mouse anti-MyHC IIb antibody (1:1000, BF-F3, DSHB, United States), and rabbit anti-β-actin antibody (1:5000; AP0060, Bioworld, China).

Techniques: Western Blot, Expressing, Control, Quantitative RT-PCR