Fig 1: Musclin predicts outcome in patients undergoing transcatheter aortic valve implantation. A, Distribution of musclin levels within the study cohort with low (<2.862 ng/mL) and high (=2.862 ng/mL) circulating musclin determined by classification and regression tree analysis. B through D, Survival curves based on (B) circulating musclin, (C) European System for Cardiac Operative Risk Evaluation (EuroSCORE) II, and (D) the combination of systemic musclin and EuroSCORE II (low risk <4% and intermediate risk =4%). HR indicates hazard ratio.
Fig 2: Elevation of muscle-secreted Musclin exacerbates HFD-induced obesity and metabolic dysfunction.MCK-Musclin and WT male mice were fed with HFD beginning at 3 months old. a Body weight of MCK-Musclin and WT mice housed at 16°C. WT vs MCK-Musclin, n = 8 vs 7. b Fasting blood glucose. WT vs MCK-Musclin, n = 11 vs 6. c Plasma insulin concentration of mice housed at 16°C. WT vs MCK-Musclin, n = 7 vs 6. d Glucose tolerance test (left) and the area under curve (AUC, right). WT vs MCK-Musclin, n = 9 vs 5. e Insulin tolerance test (left) and AUC (right) of mice housed at 16°C. WT vs MCK-Musclin, n = 9 vs 7. f Lean mass and fat mass after HFD feeding for 2 months at 16°C. WT vs MCK-Musclin, n = 6 vs 5. g Weight of indicated tissues from mice housed at 16°C. WT vs MCK-Musclin, n = 7 vs 6. h Representative H&E staining images of iWAT, eWAT, and BAT (WT vs MCK-Musclin, n = 4 vs 5). i The cell size of iWAT (n = 5 per group, left panel) and total lipid droplet area per view in BAT H&E sections (n = 7 per group, right panel) as shown in (h). a–g, i data represent mean ± SEM; n represents biologically independent animals. a Two-way ANOVA with Sidak’s multiple comparisons. b–g, i Two-tailed unpaired Student’s t-test. Experiments in a–g were repeated independently three times, and experiments in h and i were repeated independently twice with similar results. Source data are provided as a Source Data file.
Fig 3: Transgenic overexpression of Musclin inhibits adipose tissue thermogenesis.All mice used here are male and fed with chow diet. a qPCR analysis of Musclin mRNA in muscles from indicated mice (3 months old). Room temperature (RT) vs 30°C (1 week), n = 6; RT vs Cold, n = 5 vs 4. b Immunoblots of muscle lysates (left, 3 months old) and quantification of Musclin protein levels (right). n = 3. c Immunoblots of muscle lysates (left, 3.5 months old), and quantification of Musclin protein levels (right). RT vs Cold, n = 5 vs 6. d Plasma Musclin levels. RT vs 30°C, n = 6 (3 months old); RT vs Cold, n = 11 (5 months old). e Binding assay on mouse tissue sections using SEAP or SEAP-Musclin fusion proteins. SEAP, secreted alkaline phosphatase; iWAT, inguinal white adipose tissue; eWAT, epidydimal white adipose tissue. Scale bar: 100 µm. f Immunoblots of muscle lysates (left), and quantification of Musclin protein levels (right). WT vs MCK-Musclin, n = 5 vs 7. MCK, muscle creatine kinase. g RIA of plasma Musclin levels. WT vs MCK-Musclin, n = 8 vs 7. h Body weight (6 months old). WT vs MCK-Musclin, n = 6 vs 8. i iWAT and BAT weight (7 months old). WT vs MCK-Musclin, n = 5 vs 7. j Representative images of iWAT (7 months old). k Core body temperature before and 6 h following cold at 4°C (2.5 months old), n = 8. l Ratio of homeothermic mice during cold at 4°C (2.5 months old) (Homeothermia is defined as the ratio of mice with a body temperature above 31°C), n = 8. m Core temperature during cold at 8°C (7 months old). WT vs MCK-Musclin, n = 7 vs 6. n Representative infrared images (left) and the average body surface temperature (right) of mice following 3 h of CL316,243 (CL) injection (4.5 months old). n = 5. o Blood glucose following cold at 8 °C for 2 days (6.5 months old). WT vs MCK-Musclin, n = 6 vs 8. p Plasma insulin levels (5 months old). WT vs MCK-Musclin, n = 5 vs 6. Data represent mean ± SEM (n represents biologically independent animals). a–d, f–i, k, n–p Two-tailed unpaired Student’s t-test; m two-way ANOVA with Sidak’s multiple comparisons. All experiments here were repeated independently three times with similar results. Source data are provided as a Source Data file.
Fig 4: Musclin inhibits the activation of cardiac fibroblasts through NPR-B dependent activation of protein kinase G (PKG).a Measurement of cardiac fibroblast proliferation by BrDU incorporation ELISA with addition of CNP and/or Musclin (n = 5/condition, **p = 0.0039 for vehicle vs. CNP, **p = 0.0019 for vehicle vs. musclin and ***p = 0.0002). b Assessment of cardiac fibroblast migration through detection of recovery of a scratch wound after 4 h during stimulation as indicated (n = 6/condition, **p = 0.0023, ***p = 0.0003 for vehicle-treated cells vs. stimulated with Musclin, and ***p = 0.0005 vs. cells stimulated with CNP and Musclin). c–e Measurement of cardiac fibroblast proliferation by BrDU incorporation ELISA under the indicated conditions (c–e) (n = 6/condition, except siNPR2 treatment n = 7); *p = 0.0103 for siControl cells, vehicle treated vs. stimulated with CNP, **p = 0.0016 vs. stimulated with Musclin and ***p = 0.0001 vs. stimulated with CNP and Musclin, *p = 0.0103 for siNPR1 cells, vehicle treated vs. stimulated with CNP, ****p < 0.0001 vs. stimulated with Musclin and vs. stimulated with CNP and Musclin (c); *p = 0.0243, **p = 0.0055 and ***p = 0.0003 (d); **p = 0.0021 for siControl cells, vehicle treated vs. stimulated with CNP, ***p = 0.0002 vs. stimulated with Musclin, and ****p < 0.0001 vs. treated with both CNP and Musclin, ***p = 0.0002 for siNPR3 cells, vehicle treated vs. stimulated with CNP, ****p < 0.0001 vs. treated with Musclin and vs. cells treated with both CNP and Musclin (e). Assessment of cardiac fibroblast migration by scratch assay in cardiac fibroblasts (n = 6/condition) treated with siRNA, CNP and Musclin as indicated (f–h), *p = 0.036 for siControl cells, vehicle treated vs. treated with CNP, ***p < 0.0001 vs. Musclin and vs. stimulated with CNP and Musclin, *p = 0.046 for siNPR1 cells, vehicle treated vs. stimulated with CNP, **p = 0.0013 vs. Musclin and ****p < 0.0001 vs. cells treated with both CNP and Musclin (f); *p = 0.0148 for siControl cells, vehicle treated vs. treated with CNP, **p = 0.0013, and *p = 0.0148 between siControl and siNPR2 cells treated with both CNP and Musclin (g); *p = 0.0162 for siControl cells, vehicle treated vs. cells stimulated with CNP, **p = 0.0042 vs. cells treated with Musclin and ***p = 0.0004 vs. cells stimulated with both CNP and Musclin, *p = 0.0162 for siNPR3 cells, vehicle treated vs. stimulated with CNP or Musclin and ***p = 0.0002 vs. stimulated with Musclin and CNP, ***p = 0.0005 for siControl cells treated with Musclin und CNP vs. siNPR3 cells stimulated with Musclin and CNP, ***p = 0.0008 for vehicle-treated siControl cells vs. vehicle-treated siNPR3 cells (h). i, j Assessment of cardiac fibroblast proliferation and migration after stimulation with Musclin, CNP or the PKG inhibitor DT3 as indicated (for proliferation n = 7/condition, **p = 0.0018 for vehicle vs. Musclin treated cells and *p = 0.016 for cells stimulated with Musclin vs. cells treated with Musclin and DT3 (i), and for migration n = 4/condition, **p = 0.0043 for vehicle-treated cells vs. stimulated with Musclin, ***p = 0.0005 for cells treated with Musclin vs. treated with DT3 (j)). k Immunoblot from cardiac fibroblasts’ protein lysate for the indicated proteins after stimulation as indicated. GAPDH was used as loading control. The size of the proteins is indicated in kDa. Data in bar graphs are shown as mean ± standard error of the mean (SEM). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 determined by one-way ANOVA followed by the Holms–Sidak’s multiple comparisons test. Mu stands for Musclin. Source data are provided as a source data file.
Fig 5: Muscle OSTN mRNA and Musclin serum levels are reduced in patients with heart failure.a OSTN mRNA levels in vastus lateralis muscle biopsy samples from control individuals (n = 9) and from patients with sarcopenia (n = 6) or cachexia (n = 5), *p = 0.0236. b Concentrations of Musclin protein in the serum of healthy control individuals (n = 56) and of patients with severe aortic stenosis (n = 26), *p = 0.0403 (c) and of the same patients excluding the ones with diabetes mellitus (n = 18), **p = 0.0044. Data are shown as mean ± standard error of the mean (SEM). *p < 0.05, **p < 0.01 as determined by ANOVA followed by the Holms–Sidak’s multiple comparisons test (a) and the two-tailed Mann–Whitney test (b, c). Source data are provided as a source data file.
Supplier Page from CUSABIO Technology LLC for Human osteocrin,Ostn ELISA Kit