Journal: Aging Cell

Article Title: Diosgenin Alleviates Age‐Related Sarcopenia by Promoting Satellite Cell Proliferation and Myogenic Differentiation via Activation of the SIRT1 / PGC ‐1α Signaling Pathway

doi: 10.1111/acel.70651

Figure Lengend Snippet: Diosgenin promoted proliferation and myogenic differentiation of senescent satellite cells. (A) Representative images of Pax7/Ki67 double IF staining in skeletal muscle tissues, with quantitative analysis. Green: Pax7 (satellite cell marker); red: Ki67 (proliferation marker); blue: DAPI (nuclei); yellow: Pax7 + /Ki67 + double‐positive cells. Scale bar = 20 μm. (B) Cytotoxicity of diosgenin in differentiated myotubes (determination of safe concentration range, n = 3). (C) Proliferative activity of diosgenin in undifferentiated C2C12 myoblasts in the absence (left panel) or presence (right panel) of 100 μM D‐gal ( n = 3). (D) Representative WB images and quantitative analysis of the myogenic differentiation markers Myf5, Pax7, and MyHC II in mouse skeletal muscle. GAPDH was used as the internal control. (E) Representative WB images and quantitative analysis of the myogenic differentiation markers Myf5, Pax7, and MyHC II in C2C12 cells. GAPDH was used as the internal control. Data are presented as mean ± SEM. * p ‐value < 0.05, ** p ‐value < 0.01, *** p ‐value < 0.001 versus the YC group; # p ‐value < 0.05, ## p ‐value < 0.01, ### p ‐value < 0.001 versus the OC group. Statistical analysis was performed using one‐way ANOVA followed by Tukey's multiple comparisons test. All experiments were performed with at least three independent replicates.

Article Snippet: Membranes were blocked with 5% skim milk at room temperature for 1 h and incubated with the following primary antibodies at 4°C overnight: SIRT1 (1:1000; Affinity Biosciences, Liyang, China; Cat. #DF6033), PGC‐1α (1:1000; ABclonal, Wuhan, China; Cat. #AP20542C), p53 (1:1000; ABclonal, Wuhan, China; Cat. #A21630), p21 (1:1000; ABclonal, Wuhan, China; Cat. #A1483), Myf5 (1:1000; ABclonal, Wuhan, China; Cat. #A116227), MyHC II (1:1000; ABclonal, Wuhan, China; Cat. #A15293), Pax7 (1:1000; ABclonal, Wuhan, China; Cat. #A7335), and GAPDH (1:20,000; ABclonal, Wuhan, China; Cat. #AC033).

Techniques: Cell Characterization, Staining, Marker, Concentration Assay, Activity Assay, Control

Journal: Aging Cell

Article Title: Diosgenin Alleviates Age‐Related Sarcopenia by Promoting Satellite Cell Proliferation and Myogenic Differentiation via Activation of the SIRT1 / PGC ‐1α Signaling Pathway

doi: 10.1111/acel.70651

Figure Lengend Snippet: Diosgenin regulated SIRT1 to ameliorate age‐related sarcopenia. (A) Representative WB images and quantitative analysis of SIRT1 in C2C12 cells. (B) Quantitative analysis of PGC‐1α protein expression and PGC‐1α acetylation levels in C2C12 cells. (C) mRNA levels of Cpt1b in C2C12 cells. (D) mRNA levels of Metrnl and Igf1 in C2C12 cells. (E) Protein concentrations of METRNL and IGF‐1 in C2C12 cells. (F) Representative WB images and quantitative analysis of the myogenic differentiation markers Myf5, Pax7, and MyHC II in C2C12 cells. (G) Representative WB images and quantitative analysis of the senescence markers p53 and p21 in C2C12 cells. (H) Representative SA‐β‐gal staining images of C2C12 cells at 100× and 400× magnification, with quantitative analysis. (I) Molecular docking results of diosgenin with the SIRT1 protein. (J) Schematic diagram of the interactions between diosgenin and key residues of SIRT1. (K) Representative WB images reflecting the thermal stability of SIRT1 protein under different temperature gradients and the corresponding thermal stability curve; the y‐axis represents the remaining SIRT1 protein level relative to 37°C. Data are presented as mean ± SEM. * p‐ value < 0.05, ** p ‐value < 0.01, *** p ‐value < 0.001 versus the YC group; # p ‐value < 0.05, ## p ‐value < 0.01, ### p ‐value < 0.001 versus the OC group. Statistical analysis was performed using one‐way ANOVA followed by Tukey's multiple comparisons test. All experiments were performed with at least three independent replicates.

Article Snippet: Membranes were blocked with 5% skim milk at room temperature for 1 h and incubated with the following primary antibodies at 4°C overnight: SIRT1 (1:1000; Affinity Biosciences, Liyang, China; Cat. #DF6033), PGC‐1α (1:1000; ABclonal, Wuhan, China; Cat. #AP20542C), p53 (1:1000; ABclonal, Wuhan, China; Cat. #A21630), p21 (1:1000; ABclonal, Wuhan, China; Cat. #A1483), Myf5 (1:1000; ABclonal, Wuhan, China; Cat. #A116227), MyHC II (1:1000; ABclonal, Wuhan, China; Cat. #A15293), Pax7 (1:1000; ABclonal, Wuhan, China; Cat. #A7335), and GAPDH (1:20,000; ABclonal, Wuhan, China; Cat. #AC033).

Techniques: Expressing, Cell Characterization, Staining

Journal: PCN Reports: Psychiatry and Clinical Neurosciences

Article Title: Dystrophinopathy with a DMD exon 49–50 deletion in a female patient who developed schizophrenia: An autopsy case

doi: 10.1002/pcn5.70327

Figure Lengend Snippet: Microscopic findings. Klüver–Barrera staining shows some neurons densely arranged in a line across layers IV–V in parts of the visual cortex (a, b). Klüver–Barrera staining shows single‐neuronal heterotopia observed in the subependymal regions around the lateral ventricles (c, as indicated by the arrow) and in the white matter of the middle frontal gyrus (d). Anterior horn neurons of the spinal cord (e) and Betz cells in the precentral gyrus (f) are preserved. The three layers of the cerebellum and Purkinje cells are well‐preserved (g). Several eosinophilic structures are found in the gracile nucleus (h) with negative immunostaining of anti‐glial fibrillary acidic protein (i) and anti‐phosphorylated neurofilament (j). Neurofibrillary tangles and neuropil threads are observed in a limited region of the transentorhinal cortex (k). Skeletal muscle fibers display variability in size, with some showing a rounded morphology (l). Interstitial spreading and fibrosis of the myocardial fibers are observed (m). Scale bars: 100 μm (e, g, m), 50 μm (a, c, f, h–l), and 20 μm (b, d).

Article Snippet: Anti‐phosphorylated neurofilament , Covance , Monoclonal , Mouse , 1:5000 , Heat antigen retrieval.

Techniques: Staining, Immunostaining