pgc1α antibodies  (Novus Biologicals)

Journal: Food Science & Nutrition

Article Title: Mitigating Dexamethasone‐Induced Muscle Wasting and Mitochondrial Impairment in Mice on a High‐Fat and High‐Sucrose Diet With Peanut Sprout Extract

doi: 10.1002/fsn3.71469

Figure Lengend Snippet: Peanut sprout extract (PSE) facilitated mitochondrial activation in the skeletal muscle of Dexamethasone (Dex) and high‐fat and high‐sucrose (HFHS)‐treated mice. Male C57BL/6 mice are fed a control diet (Control, n = 5) or HFHS diet with daily administration of saline or PSE (10 mg/kg body weight [BW]) for 10 weeks. Dex was injected during the last six days to induce muscle atrophy (Dex + HFHS, n = 8; Dex + HFHS + PSE, n = 9 per group). (A) Western blot analysis of peroxisome proliferator‐activated receptor‐γ coactivator 1‐α (PGC1α) and mitochondrial transcription factor A (TFAM) in gastrocnemius muscle and relative protein expression. GAPDH served as a control. (B) Western blot of oxidative phosphorylation (OXPHOS) protein expression in gastrocnemius muscle and relative protein expression. GAPDH served as a control. Dashed lines in the western blot images indicate boundaries between different experimental groups (Dex + HFHS vs. Dex + HFHS + PSE), while all samples were processed on the same blot. Data are expressed as the mean ± standard error of the mean (SEM); n.s., not significant; exact p ‐values were shown in the graph by comparing Control vs. Dex + HFHS or Dex + HFHS vs. Dex + HFHS + PSE by Student's t ‐test.

Article Snippet: Subsequently, they were incubated overnight at 4°C with primary antibodies against Muscle atrophy F‐box (MAFbx/Atrogin‐1), Muscle specific RING finger protein (MuRF1) (Santa Cruz Biotechnology, CA, USA); glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH); oxidative phosphorylation (OXPHOS) (Abcam, Cambridge, MA, USA); peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha (PGC1α) (Novus Biologicals LLC); Mitochondrial transcription factor A (TFAM) (Abcam, Cambridge, MA, USA), and total or phospho nuclear factor‐κB (NF‐κB) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Activation Assay, Control, Saline, Injection, Western Blot, Expressing, Phospho-proteomics

Journal: Food Science & Nutrition

Article Title: Mitigating Dexamethasone‐Induced Muscle Wasting and Mitochondrial Impairment in Mice on a High‐Fat and High‐Sucrose Diet With Peanut Sprout Extract

doi: 10.1002/fsn3.71469

Figure Lengend Snippet: Effects of PSE on Dex‐induced muscle atrophy and mitochondrial function in C2C12 cells. The C2C12 cells were incubated with PSE extract at different doses (0–200 μg/mL). XTT reagent was added for 3 h into 96‐well plates to measure cell viability. (A) cell viability by XTT. (B) The C2C12 cells were incubated with 25 μg/mL of PSE for five days during differentiation. After starving for 1 h, they are treated with 10 μM Dex and harvested after 24 h. Atrophy and inflammatory gene expressions of Atrogin‐1 , Murf1, Il1β, Tnfα , and Pgc1α a re quantified using real‐time PCR. (C) Oxygen consumption rate (OCR) in C2C12 cells treated with Veh (black circle, dot line), Dex (transparent square, black line) or Dex + PSE (Gray triangle, gray line) as determined by Seahorse extracellular analyzer. The C2C12 cells are treated with 25 μg/mL PSE for five days during differentiation. After starving for 1 h, they are treated with 10 μM Dex. Arrow indicates the addition of respiratory inhibitors of oligomycin (Oligo), Carbonyl cyanide 4‐trifluoromethoxyphenylhydrazone (FCCP) and combination of antimycin A and rotenone (Rot/AA). (D) OCR profiles. Data are expressed as the mean ± standard error of the mean (SEM); Bars with different letters are significantly different by one‐way ANOVA with Duncan or Tukey's comparison test. n.s., not significant; exact p ‐value were shown in graph by comparing Control vs. Dex + HFHS or Dex + HFHS vs. Dex + HFHS+PSE by Student's t ‐test.

Article Snippet: Subsequently, they were incubated overnight at 4°C with primary antibodies against Muscle atrophy F‐box (MAFbx/Atrogin‐1), Muscle specific RING finger protein (MuRF1) (Santa Cruz Biotechnology, CA, USA); glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH); oxidative phosphorylation (OXPHOS) (Abcam, Cambridge, MA, USA); peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha (PGC1α) (Novus Biologicals LLC); Mitochondrial transcription factor A (TFAM) (Abcam, Cambridge, MA, USA), and total or phospho nuclear factor‐κB (NF‐κB) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Incubation, Real-time Polymerase Chain Reaction, Comparison, Control