follistatin  (MedChemExpress)

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Fibro‐Adipogenic Progenitors Regulate Orofacial Neuromuscular Junction Regeneration via Myostatin

doi: 10.1002/jcsm.70264

Figure Lengend Snippet: Recombinant MSTN suppressed AChR clustering and miR‐206 expression. (a) AChR and MyHC staining of MuSCs cultures treated with or without recombinant MSTN (100 ng/mL) and follistatin (500 ng/mL). Scale bar = 50μm. (b) Graphs show (from left to right) the quantification of differentiation index, myotube area, AChR cluster counts normalized to myotube area and ratio of AChR/myotube area. n = 3. (c) The levels of miR‐206 expression in MuSCs cultures. Mature myotubes differentiated from MAS‐derived MuSCs were treated with control, agrin alone, agrin plus MSTN or agrin plus MSTN and follistatin. (d) AChR and MyHC staining of myotubes transfected with miR‐206 mimics or negative control (NC) prior to treatment with agrin and MSTN. Scale bar = 50 μm. (e) Graphs show (from left to right) the quantification of differentiation index, myotube area, AChR cluster counts normalized to myotube area and ratio of AChR/myotube area. n = 3. The data are shown as mean ± SD. One‐way ANOVA followed by Tukey's post hoc test was used. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mature myotubes were treated with 100 ng/mL agrin (HY‐ P79236 , MedChemExpress, China) in DM or conditioned medium (CM) of 7 dpi FAPs supplemented with 2% HS for 16 h. For recombinant MSTN treatment, 100 ng/mL MSTN (HY‐ P72632 , MedChemExpress, China) with or without 500 ng/mL follistatin (HY‐ P70315 , MedChemExpress, China) was administered together with agrin treatment.

Techniques: Recombinant, Expressing, Staining, Derivative Assay, Control, Transfection, Negative Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Activin A–Endothelin-1 Axis Governs Pulmonary Vascular Remodeling: Mechanistic Basis for Emerging Therapies in PAH

doi: 10.1161/ATVBAHA.125.323681

Figure Lengend Snippet: Activin A induces excessive ET-1 (endothelin-1) production in pulmonary artery endothelial cells (PAECs), reversible by follistatin or bosentan. A and B , Quantitative real-time polymerase chain reaction analysis of ET-1 mRNA expression ( A , n=6 biologically independent samples per group) and ELISA measurement of ET-1 concentration in culture medium ( B , n=3–4) from PAECs treated for 6 hours with recombinant activin A (100 ng/mL) or vehicle. C and D , ET-1 mRNA expression ( C , n=6) and secreted ET-1 concentration ( D , n=4-5) in PAECs 48 hours after INHBA (inhibin β-A) overexpression (OE) or GFP (green fluorescent protein) control retroviral transfection. E and F , INHBA mRNA expression ( E , n=6) and activin A concentration in culture medium ( F , n=4) in PAECs treated for 6 hours with recombinant ET-1 (100 nmol/L) or vehicle. G , ET-1 mRNA expression in PAECs treated for 6 hours with vehicle or recombinant activin A in the presence of vehicle (VEH), FST (follistatin; 100 ng/mL), bosentan (BOS; 10 μM), or both (FST+BOS; n=4). H , ET-1 mRNA expression in PAECs 48 hours after INHBA OE or GFP transfection, followed by 24 hours of treatment with VEH, FST (100 ng/mL), BOS (10 μM), or FST+BOS (n=4). I , ET-1 mRNA expression in PAECs treated for 6 hours with vehicle or recombinant activin A in the presence of VEH or ACTRIIA-Fc (activin receptor type IIa fusion protein; 2500 ng/mL; n=3-4). J , ET-1 mRNA expression in PAECs 48 hours after INHBA OE or GFP transfection, followed by 24 hours of treatment with VEH or ACTRIIA-Fc (2500 ng/mL; n=4). Data are mean±SEM. P <0.05 is deemed statistically significant. Statistical tests: 2-sided Student t test for A through F ; 1-way ANOVA with Tukey post hoc test for G and H .

Article Snippet: For indicated experiments, PAECs were treated for 24 hours with FST (100 ng/mL; no. 4889-FN-025; R&D Systems), bosentan (10 μM; no. SML1265; Sigma-Aldrich), ALK (activin receptor–like kinase) 4/5/7 inhibitor SB505124 (5 μM; no. S2186; Selleck), ALK1/2/6 inhibitor K02288 (1 μM; no. S7359; Selleck), the activin receptor type IIA fusion protein ACTRIIA-Fc (activin receptor type IIA fusion protein; 2500 ng/mL), or vehicle.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Recombinant, Over Expression, Control, Retroviral, Transfection

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Activin A–Endothelin-1 Axis Governs Pulmonary Vascular Remodeling: Mechanistic Basis for Emerging Therapies in PAH

doi: 10.1161/ATVBAHA.125.323681

Figure Lengend Snippet: Activin A–driven ET-1 (endothelin-1) contributes to pulmonary artery endothelial cell (PAEC) dysfunction. A and B , Representative images ( A ) and quantification ( B ) of chord length and number of branching points in a Matrigel tube formation assay (n=3–4) using PAECs transfected with GFP (green fluorescent protein) or INHBA (inhibin β-A) overexpression (OE), treated with vehicle (VEH), FST (follistatin; 100 ng/mL), bosentan (BOS; 10 μM), or both (FST+BOS). C and D , Representative images ( C ) and quantification ( D ) of apoptotic cells assessed by TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) staining (n=3–4) under serum starvation. Apoptotic cells are indicated by white arrows. E , Cell proliferation measured by WST-1 (water-soluble tetrazolium-1) assay in GFP- or INHBA OE–transfected PAECs treated with VEH, FST, BOS, or FST+BOS (n=3). Data are mean±SEM. P <0.05 is deemed statistically significant. Statistical test: 1-way ANOVA with Tukey post hoc test for B , D , and E . AU indicates a bsorbance units; and DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: For indicated experiments, PAECs were treated for 24 hours with FST (100 ng/mL; no. 4889-FN-025; R&D Systems), bosentan (10 μM; no. SML1265; Sigma-Aldrich), ALK (activin receptor–like kinase) 4/5/7 inhibitor SB505124 (5 μM; no. S2186; Selleck), ALK1/2/6 inhibitor K02288 (1 μM; no. S7359; Selleck), the activin receptor type IIA fusion protein ACTRIIA-Fc (activin receptor type IIA fusion protein; 2500 ng/mL), or vehicle.

Techniques: Tube Formation Assay, Transfection, Over Expression, TUNEL Assay, Staining

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Activin A–Endothelin-1 Axis Governs Pulmonary Vascular Remodeling: Mechanistic Basis for Emerging Therapies in PAH

doi: 10.1161/ATVBAHA.125.323681

Figure Lengend Snippet: Endothelial cell (EC)–derived activin A–induced ET-1 (endothelin-1) alters vasoconstrictive properties of the pulmonary vasculature. A , eNOS (endothelial NO synthase) mRNA expression in pulmonary artery endothelial cells (PAECs) 48 hours after GFP (green fluorescent protein) or INHBA (inhibin β-A) overexpression (OE), treated with vehicle (VEH), FST (follistatin; 100 ng/mL), bosentan (BOS; 10 μM), or FST+BOS for 24 hours (n=3). B , Representative immunoblots and quantification of p-eNOS (phosphorylated eNOS) and total eNOS under the same conditions (n=3–4). C , ET-1 concentration in pulmonary artery smooth muscle cell (PASMC) culture medium after coculture with PAECs treated with recombinant activin A for 24 or 48 hours (n=3). D , ET-1 and INHBA mRNA expression in PASMCs cocultured for 48 hours with GFP- or INHBA OE–PAECs, with VEH, FST, BOS, or FST+BOS added for the last 24 hours (n=3–4). E , PASMC mRNA expression of PCNA (proliferating cell nuclear antigen), fibronectin, SM22α (smooth muscle protein 22-α), and α-SMA (α-smooth muscle actin) under the same conditions (n=3–4). F , Representative immunoblots and quantification of MYH11 (myosin heavy chain 11), SM22α, and MMP2 (matrix metalloproteinase-2) in PASMCs cocultured with GFP- or INHBA OE–PAECs, treated as in E (n=3). Data are mean±SEM. P <0.05 is deemed statistically significant. Tests: 2-sided Student t test for C ; 1-way ANOVA with Tukey post hoc test for A , B , D through F .

Article Snippet: For indicated experiments, PAECs were treated for 24 hours with FST (100 ng/mL; no. 4889-FN-025; R&D Systems), bosentan (10 μM; no. SML1265; Sigma-Aldrich), ALK (activin receptor–like kinase) 4/5/7 inhibitor SB505124 (5 μM; no. S2186; Selleck), ALK1/2/6 inhibitor K02288 (1 μM; no. S7359; Selleck), the activin receptor type IIA fusion protein ACTRIIA-Fc (activin receptor type IIA fusion protein; 2500 ng/mL), or vehicle.

Techniques: Derivative Assay, Expressing, Over Expression, Western Blot, Concentration Assay, Recombinant

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Activin A–Endothelin-1 Axis Governs Pulmonary Vascular Remodeling: Mechanistic Basis for Emerging Therapies in PAH

doi: 10.1161/ATVBAHA.125.323681

Figure Lengend Snippet: Activin A–derived ET-1 (endothelin-1) drives multiple proremodeling pathways in pulmonary artery endothelial cells (PAECs). A , mRNA expression of BMP4 (bone morphogenetic protein 4), SLUG (snail family transcriptional repressor 2), SNAIL (snail family transcriptional repressor 1), VE-cadherin (vascular endothelial cadherin), fibronectin, and SOD2 (superoxide dismutase 2) in GFP (green fluorescent protein)- or INHBA (inhibin β-A) overexpression (OE)–PAECs treated with vehicle (VEH), FST (follistatin), bosentan (BOS), or FST+BOS for 24 hours (n=3–4). B , Representative immunoblots and quantification of vimentin, SOD2, and NRF2 (nuclear factor, erythroid 2-related factor 2) under the same conditions (n=3). C , Reactive oxygen species (ROS) production in GFP- or INHBA OE–PAECs treated as in A , measured at 4, 6, 16, and 24 hours (n=3–4). Data are mean±SEM. P <0.05 is deemed statistically significant. Tests: 1-way ANOVA with Tukey post hoc test for A and B ; 2-way ANOVA with Tukey post hoc test for C .

Article Snippet: For indicated experiments, PAECs were treated for 24 hours with FST (100 ng/mL; no. 4889-FN-025; R&D Systems), bosentan (10 μM; no. SML1265; Sigma-Aldrich), ALK (activin receptor–like kinase) 4/5/7 inhibitor SB505124 (5 μM; no. S2186; Selleck), ALK1/2/6 inhibitor K02288 (1 μM; no. S7359; Selleck), the activin receptor type IIA fusion protein ACTRIIA-Fc (activin receptor type IIA fusion protein; 2500 ng/mL), or vehicle.

Techniques: Derivative Assay, Expressing, Over Expression, Western Blot

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Activin A–Endothelin-1 Axis Governs Pulmonary Vascular Remodeling: Mechanistic Basis for Emerging Therapies in PAH

doi: 10.1161/ATVBAHA.125.323681

Figure Lengend Snippet: Canonical SMAD2/3 signaling mediates activin A–induced ET-1 (endothelin-1) expression. A , Representative immunoblots and quantification of p-SMAD2/3 (phosphorylated SMAD2/3; small mother against decapentaplegic family member 2/3) and total SMAD2/3 in GFP (green fluorescent protein)- or INHBA (inhibin β-A) overexpression (OE)–pulmonary artery endothelial cells (PAECs) treated with vehicle (VEH), FST (follistatin), bosentan (BOS), or FST+BOS for 24 hours (n=3). B , Immunoblots showing p-SMAD2/3 and total SMAD2/3 in GFP- or INHBA OE–PAECs treated with SB505124 (5 μM) or vehicle for 24 hours. C , ET-1 mRNA expression in GFP- or INHBA OE–PAECs treated with SB505124 or vehicle for 24 hours ( left , n=3) and in PAECs treated for 6 hours with activin A±SB505124 ( right , n=3–4). D , Immunoblots showing SMAD2/3 and β-actin in PAECs pretreated with SMAD2 siRNA (small interfering RNA; siSMAD2), SMAD3 siRNA (siSMAD3), dual SMAD2-SMAD3 siRNA (dual siSMAD), or control siRNA (siNC [siRNA negative control]). E , ET-1 mRNA expression in GFP- or INHBA OE–PAECs pretreated with siSMAD2, siSMAD3, dual siSMAD, or siNC (n=3–4). F , INHBA and ET-1 mRNA expression in PAECs exposed to normoxia or hypoxia (0.1% O 2 , 24 hours; n=6). G and H , ET-1 mRNA expression in PAECs under hypoxia treated with FST ( G ) or SB505124 ( H ; n=3). Data are mean±SEM. P <0.05 is deemed statistically significant. Tests: 1-way ANOVA with Tukey post hoc test for A , C , E , G , and H ; 2-sided Student t test for F .

Article Snippet: For indicated experiments, PAECs were treated for 24 hours with FST (100 ng/mL; no. 4889-FN-025; R&D Systems), bosentan (10 μM; no. SML1265; Sigma-Aldrich), ALK (activin receptor–like kinase) 4/5/7 inhibitor SB505124 (5 μM; no. S2186; Selleck), ALK1/2/6 inhibitor K02288 (1 μM; no. S7359; Selleck), the activin receptor type IIA fusion protein ACTRIIA-Fc (activin receptor type IIA fusion protein; 2500 ng/mL), or vehicle.

Techniques: Expressing, Western Blot, Over Expression, Small Interfering RNA, Control, Negative Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Activin A–Endothelin-1 Axis Governs Pulmonary Vascular Remodeling: Mechanistic Basis for Emerging Therapies in PAH

doi: 10.1161/ATVBAHA.125.323681

Figure Lengend Snippet: In vivo activin A inhibition improves pulmonary hypertension (PH) phenotype comparably or more than ET-1 (endothelin-1) blockade. A , Experimental design: wild-type (WT) and VE-cadherin (vascular endothelial cadherin)–INHBA (inhibin β-A)-Tg (TG/transgenic) mice were exposed to hypoxia (10% O 2 ) for 3 weeks, with vehicle (VEH), FST (follistatin; 8.5 μg/kg), bosentan (BOS; 30 mg/kg), or FST+BOS administered during the final 2 weeks. B , Right ventricular systolic pressure (RVSP; n=4–9). C , Fulton index (RV/[LV+S] [right ventricle to left ventricle plus septum] ratio; n=4–8). D , Representative hematoxylin and eosin–stained lung sections. Blue arrows indicate vessels. E , Representative immunofluorescent staining of α-SMA (α-smooth muscle actin protein; green, SMC [smooth muscle cell] marker), vWF (von Willebrand Factor; red, endothelial cell [EC] marker), and DAPI (4′,6-diamidino-2-phenylindole; blue, nuclei). White arrows indicate vessels. F , Quantification of pulmonary artery muscularization (non-, partial-, full; n=12–15 fields from 3–4 mice). PA indicates pulmonary artery. G , Lung ET-1 mRNA expression (n=3–4). H , mRNA expression of INHBA, ET-1, eNOS (endothelial NO synthase), SOD2 (superoxide dismutase 2), fibronectin, SLUG (snail family transcriptional repressor 2), CD31 (cluster of differentiation 31), and BMP4 (bone morphogenetic protein 4) in lung ECs isolated from WT and TG mice (n=3–4). Data are mean±SEM. P <0.05 is deemed statistically significant. Tests: 1-way ANOVA with Tukey post hoc test for B , C , and G ; 2-way ANOVA with Tukey post hoc test for F ; 2-sided Student t test for H .

Article Snippet: For indicated experiments, PAECs were treated for 24 hours with FST (100 ng/mL; no. 4889-FN-025; R&D Systems), bosentan (10 μM; no. SML1265; Sigma-Aldrich), ALK (activin receptor–like kinase) 4/5/7 inhibitor SB505124 (5 μM; no. S2186; Selleck), ALK1/2/6 inhibitor K02288 (1 μM; no. S7359; Selleck), the activin receptor type IIA fusion protein ACTRIIA-Fc (activin receptor type IIA fusion protein; 2500 ng/mL), or vehicle.

Techniques: In Vivo, Inhibition, Transgenic Assay, Staining, Marker, Expressing, Isolation

Journal: Journal of the International Society of Sports Nutrition

Article Title: Combined resistance exercise and essential amino acid intake enhance follistatin/myostatin ratio and muscle fitness in older women: a randomized controlled trial

doi: 10.1080/15502783.2026.2646626

Figure Lengend Snippet: Alterations in the follistatin/myostatin ratio. The follistatin/myostatin ratio was determined based on pre- and post-intervention levels for each biomarker. Percentage changes in the ratio were determined using the following formula: [(Post follistatin / Post myostatin) / (Pre follistatin / Pre myostatin) − 1] × 100. Differences among groups were evaluated with one-way ANOVA followed by a Bonferroni post hoc test: * p < 0.05; *** p < 0.001. Effect sizes (η²p) were interpreted as small (0.01–0.059), medium (0.06–0.139), and large (≥0.14).

Article Snippet: Serum concentrations of fasting insulin (440132, Beckman Coulter, California, CA, USA), serum levels of myokine biomarkers including IL-15, irisin, myostatin, and follistatin (DY247, DY9420-05, DY788-05, and DY669: R&D Systems, Minneapolis, MN, USA), and inflammatory cytokine biomarkers such as IL-1β, IL-6, and tumour necrosis factor-alpha (TNF- α ) (DY201, DY206, and DY210: R&D Systems, Minneapolis, MN, USA) were measured using DuoSet TM enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems, Minneapolis, MN, USA) in accordance with the manufacturer’s instructions.

Techniques: Biomarker Discovery