Journal: The FASEB Journal

Article Title: Parthenolide Attenuates Skeletal Muscle Atrophy Through Regulation of Protein Homeostasis and Inhibition of Inflammation

doi: 10.1096/fj.202504024RR

Figure Lengend Snippet: Parthenolide promotes C2C12 myotube differentiation. (A) Schematic diagram illustrating the experimental design for the 7‐day parthenolide treatment of C2C12 myoblasts. (B) Cell viability assessment using a CCK‐8 assay following treatment with various concentrations of parthenolide in C2C12 myoblasts. (C–E) C2C12 myotubes were treated with parthenolide (0–400 nM) for 48 h. (C) Following the preparation of whole cell lysates, Western blot analysis was used to assess the expression of the early differentiation markers MyoD1 and myogenin, with GAPDH serving as a control to ensure equal protein loading. (D, E) Immunofluorescence staining for MyHC was performed. MyHC is labeled in green, and the nuclei were counterstained with DAPI (blue). Myotube diameter increased with increasing parthenolide concentration. Scale bar = 50 μm. The data are presented as the mean ± SD from three independent experiments. Statistical analysis was performed using one‐way ANOVA to compare parthenolide treatment groups (50, 100, 200, and 400 nM) with vehicle control (0 nM parthenolide) group. * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: Primary antibodies against MYOD1 (18943‐1‐AP), GAPDH (60004‐1‐Ig), and mTOR (66888‐1‐Ig) were obtained from Proteintech China (Wuhan, Hubei, China).

Techniques: CCK-8 Assay, Western Blot, Expressing, Control, Immunofluorescence, Staining, Labeling, Concentration Assay

Journal: Food Science & Nutrition

Article Title: Four Dairy Products Mitigates Sarcopenia in Mice by Modulating Muscle Inflammation, Autophagy, and Protein Degradation

doi: 10.1002/fsn3.70540

Figure Lengend Snippet: Western blot of PI3K, Akt, AMPK, mTOR, p‐PI3K, p‐Akt, p‐AMPK, p‐mTOR, MyoD1, MyoG, LC3B, p62, and Beclin1 protein levels in sarcopenic mice treated for 8 weeks with goat and bovine milk. (a) Western blot images. (b–j) Quantification of Western blot bands. # indicates significant difference compared with the NC group ( p < 0.05); * indicates significant difference compared to the SOP group ( p < 0.05).

Article Snippet: Common feedstuffs according to the national standard of China for laboratory mouse feed (GB 14924.3) were obtained from Beijing Keaoxieli Feed Co. Ltd. Dexamethasone was purchased from Beijing MREDA Technology Co. Ltd., milk powder from Hyproca Nutrition Co. Ltd., atropine sulfate monohydrate from Shanghai Macklin Biochemical Co. Ltd., and Zoletil 50 from France LDBIO Diagnostics Co. Ltd. Primary antibodies against PI3K, Akt, mammalian target of rapamycin (mTOR), adenosine 5′‐monophosphate‐activated protein kinase (AMPK), p‐PI3K, p‐Akt, p‐mTOR, p‐AMPK, LC3B, and Beclin1 were used from CST Biological Reagents Co. Ltd., primary antibody against MyoD1 from ABclonal Biotech Co. Ltd., and primary antibodies against MyoG, p62, and β‐actin and secondary antibodies from Abcam Co. Ltd.

Techniques: Western Blot

Journal: Animals : an Open Access Journal from MDPI

Article Title: Comparative Analysis of Different Extracellular Matrices for the Maintenance of Bovine Satellite Cells

doi: 10.3390/ani14233496

Figure Lengend Snippet: ( A ) Bovine satellite cells were sorted by CD29 (APC), CD31 (FITC), CD45 (FITC), and CD56 (PE−Cy7) antibodies and Hoechst. Cells were first distinguished as live or dead cells by Hoechst. CD31 and CD45 negative cells were then distinguished. Next, CD29 and CD56 double-positive cells were sorted. ( B ) The isolated CD29 + , CD56 + , CD31 − , and CD45 − , bovine satellite cells were stained with PAX7 (green), MYOD1 (red) antibody, and DAPI (blue). Scale bar indicates 100 μm. ( B ) Comparative analysis of cell proliferation of bovine satellite cells cultured on gelatin-, fibronectin-, and collagen-coated dishes for short (P2)- and long (P10)-term culture ( p < 0.001). n = 9. Each letter (a, b, c) represents significant differences. Values are presented as means ± SE. ( C ) The cell cycles of the bovine satellite cells were stained with Propidium iodide (PI). The bovine satellite cultured on gelatin-, fibronectin-, and collagen-coated dishes for short- and long-term culture were then analyzed with flow cytometry. n = 3. Each letter (a, b, c) represents significant differences ( p < 0.001). Values are presented as means ± SE.

Article Snippet: Cells were then stained overnight with primary antibodies against anti-MyoD1 (Polyclonal, 1:200, Proteintech, Rosemont, IL, USA) and Pax7 (anti-paired box 7) (Pax7 monoclonal, 1:50, DHSB, Iowa, IA, USA) at 4 °C.

Techniques: Isolation, Staining, Cell Culture, Flow Cytometry

Journal: Animals : an Open Access Journal from MDPI

Article Title: Comparative Analysis of Different Extracellular Matrices for the Maintenance of Bovine Satellite Cells

doi: 10.3390/ani14233496

Figure Lengend Snippet: ( A ) Bovine satellite cells cultured on gelatin-, fibronectin-, and collagen-coated dishes at short- and long-term culture were stained with PAX7 (green), MYOD1 (red) antibody, and DAPI (blue). Scale bar indicates 100 μm. ( B ) Comparative analysis of Pax7 + MyoD1 − , Pax7 − MyoD1 + , and Pax7 + MyoD1 + cell populations between gelatin, fibronectin, and collagen in short-term culture. Population of Pax7 + MyoD1 − ( p < 0.01) n = 3. Each letter (a, b) represents significant differences. Values are presented as means ± SE. ( C ) Comparative analysis of Pax7 + MyoD1 − , Pax7 − MyoD1 + , and Pax7 + MyoD1 + cell populations between gelatin, fibronectin, and collagen in long-term culture. Pax7 + MyoD1 − ( p < 0.005), Pax7 − MyoD1 + ( p < 0.05), Pax7 + MyoD1 + ( p < 0.05), n = 3. Each letter (a, b, c) represents significant differences. Values are presented as means ± SE.

Article Snippet: Cells were then stained overnight with primary antibodies against anti-MyoD1 (Polyclonal, 1:200, Proteintech, Rosemont, IL, USA) and Pax7 (anti-paired box 7) (Pax7 monoclonal, 1:50, DHSB, Iowa, IA, USA) at 4 °C.

Techniques: Cell Culture, Staining

Journal: Animals : an Open Access Journal from MDPI

Article Title: Comparative Analysis of Different Extracellular Matrices for the Maintenance of Bovine Satellite Cells

doi: 10.3390/ani14233496

Figure Lengend Snippet: ( A ) Gene expression levels of Pax7, Pax3, Myf5, MyoD1, and MyoG in BSCs cultured on different ECMs in short-term culture (n = 3). Significant differences are indicated by different letters (a, b, c; p values: Pax7, Pax3, Myf5 < 0.005; MyoD1, MyoG < 0.0001). Values are presented as means ± SE. ( B ) Gene expression levels of Pax7, Pax3, Myf5, MyoD1, and MyoG in BSCs cultured on different ECMs in long-term culture (n = 3). Significant differences are indicated by different letters (a, b, c; p values: Pax7, Myf5 < 0.005; Pax3 < 0.05; MyoD1, MyoG < 0.0001). Values are presented as means ± SE.

Article Snippet: Cells were then stained overnight with primary antibodies against anti-MyoD1 (Polyclonal, 1:200, Proteintech, Rosemont, IL, USA) and Pax7 (anti-paired box 7) (Pax7 monoclonal, 1:50, DHSB, Iowa, IA, USA) at 4 °C.

Techniques: Gene Expression, Cell Culture