fst  (R&D Systems)

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: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: Establishment and characterization of GC (Gemcitabine and Cisplatin)-resistant bladder cancer cell lines and identification of resistance-related proteins. ( A ) GC-resistant T24-R and UC3-R cell lines were generated by gradually increasing GC concentrations. Created in BioRender ( https://BioRender.com ). ( B ) Dose-response curves and calculated half-maximal inhibitory concentration (IC50) values for cisplatin (upper panel) and gemcitabine (lower panel) in parental (T24, UC3) and GC-resistant (T24-R, UC3-R) cell lines. Data are presented as the mean ± SD from at least three independent experiments. ( C ) Apoptosis rates of parental and resistant cell lines after treatment with cisplatin, as determined by flow cytometry. Data are presented as the mean ± SD ( n ≥ 3). ( D ) Transcriptomic and proteomic analyses identified FN1 (Fibronectin), EEF1A2 (Eukaryotic Translation Elongation Factor 1 Alpha 2), MRC2 (Mannose Receptor C-Type 2), RAB6B (RAB6B, Member RAS Oncogene Family), THBS1 (Thrombospondin 1), DYSF (Dysferlin), TMOD1 (Tropomodulin 1), NES (Nestin), and APOE (Apolipoprotein E) as overexpressed in GC-resistant cells. ( E ) RT-qPCR (Reverse Transcription Quantitative Polymerase Chain Reaction) and Western blot confirmed FN1 overexpression in T24-R and UC3-R. ( F ) FN1 staining was stronger in GC-resistant bladder cancer tissues (Chemotherapy Sensitive Group: n = 6; Chemotherapy Resistant Group: n = 6). For all panels, a t -test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Statistical significance was set at p < 0.05. ns, not significant, * p < 0.05, ** p < 0.01

Article Snippet: For exogenous FN1 stimulation, recombinant human FN1 protein (rFN1) (R&D Systems, 1030-FN, Minneapolis, MN, USA) was used.

Techniques: Generated, Concentration Assay, Flow Cytometry, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Over Expression, Staining

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: ITGB4 is critical for FN1-mediated chemotherapy resistance in bladder cancer cells. ( A ) Differential expression analysis revealed that ITGB4 (highlighted by the red box) was significantly overexpressed in T24-R cells compared to T24, suggesting a role in FN1-mediated resistance. ( B ) Structural modeling shows multiple binding sites between FN1 (blue ribbon) and ITGB4 (green ribbon). ( C ) The interaction between FN1 and ITGB4 had a binding score of −318.75 with a confidence score of 96%, indicating a stable interaction. ( D ) The Co-IP experiment confirmed that there is a mutual binding interaction between FN1 and ITGB4. ( E ) Adding rFN1 to resistant strains increased FAK (Y397) phosphorylation and inhibited apoptosis, whereas ITGB4 silencing reversed these effects, highlighting the dependency of FN1-mediated resistance on ITGB4 expression and activation. For panels ( A , C , E ), a t -test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups

Article Snippet: For exogenous FN1 stimulation, recombinant human FN1 protein (rFN1) (R&D Systems, 1030-FN, Minneapolis, MN, USA) was used.

Techniques: Quantitative Proteomics, Binding Assay, Co-Immunoprecipitation Assay, Phospho-proteomics, Expressing, Activation Assay

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: FN1 silencing enhances cisplatin-induced apoptosis in bladder cancer cells. ( A ) FN1 knockdown efficiency in T24-R and UC3-R cells was confirmed by Western blot analysis. ( B ) FN1 knockdown efficiency in T24-R and UC3-R cells was confirmed by RT-qPCR analysis. ( C ) Silencing FN1 significantly increased apoptosis in the resistant cell lines by TUNEL staining. ( D ) Silencing FN1 reduced the IC50 of cisplatin in the resistant cell lines by CCK-8 assay. ( E ) In resistant cells, FN1 knockdown induced the expression of pro-apoptotic mediators, including BAX and cleaved-caspase-3, but suppressed levels of the anti-apoptotic protein Bcl-2. ( F ) Cell apoptosis rates were quantified by flow cytometry. For all panels, a t-test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Statistical significance was set at p < 0.05. ns, not significant, * p < 0.05, ** p < 0.01

Article Snippet: For exogenous FN1 stimulation, recombinant human FN1 protein (rFN1) (R&D Systems, 1030-FN, Minneapolis, MN, USA) was used.

Techniques: Knockdown, Western Blot, Quantitative RT-PCR, TUNEL Assay, Staining, CCK-8 Assay, Expressing, Flow Cytometry

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: FN1 silencing inhibits tumor growth in vivo , enhancing cisplatin sensitivity. ( A ) Representative images of tumors from each treatment group. ( B ) Tumor volume growth curves over time measured in the T24-R xenograft model. ( C ) Apoptosis in tumor tissues was detected by TUNEL staining. ( D ) Immunohistochemical analysis showed decreased FN1 expression in tumor sections from the FN1 knockdown group. For all panels, a t -test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Statistical significance was set at p < 0.05. * p < 0.05, *** p < 0.001

Article Snippet: For exogenous FN1 stimulation, recombinant human FN1 protein (rFN1) (R&D Systems, 1030-FN, Minneapolis, MN, USA) was used.

Techniques: In Vivo, TUNEL Assay, Staining, Immunohistochemical staining, Expressing, Knockdown

Journal: Oncology Research

Article Title: The FN1-ITGB4 Axis Drives Acquired Chemoresistance in Bladder Cancer by Activating FAK Signaling

doi: 10.32604/or.2025.072084

Figure Lengend Snippet: FN1 regulates FAK (Y397) phosphorylation and mediates cisplatin resistance in bladder cancer cells. ( A ) Silencing FN1 in resistant cell lines reduced the phosphorylation of FAK (Y397) as detected by Western blot. ( B ) T24 and UC3 parental and resistant cells were treated with a fixed dose of cisplatin along with a gradient of rFN1 for 48 h. Phosphorylation of FAK (Y397) was assessed by Western blot. Resistant cells (T24-R, UC3-R) showed sensitivity to rFN1 at lower concentrations under cisplatin stress. ( C ) Silencing FN1 in resistant cell lines reduced the phosphorylation of FAK (Y397) as detected by immunofluorescence. ( D ) rFN1 addition increased resistance index in resistant strains. ( E ) The addition of rFN1 reduced apoptosis in resistant strains. For all panels, a t -test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. Statistical significance was set at p < 0.05. ns, not significant, * p < 0.05, *** p < 0.001

Article Snippet: For exogenous FN1 stimulation, recombinant human FN1 protein (rFN1) (R&D Systems, 1030-FN, Minneapolis, MN, USA) was used.

Techniques: Phospho-proteomics, Western Blot, Immunofluorescence

Journal: iScience

Article Title: A tumor-microvessel on-a-chip reveals a mechanism for cancer cell cluster intravasation

doi: 10.1016/j.isci.2025.112517

Figure Lengend Snippet: Activin inhibition reduced metastatic cell behaviors with minimal impact on EndoMT (A) Schematic illustration of the concept of in vitro semi-3D experiment treated with follistatin. Follistatin neutralizes activin secreted by HUVECs or AKTP organoids. (B) Schematic illustration of the in vitro semi-3D experiment. 0.1% BSA in PBS as a vehicle was added to the control. Image acquisition and expression analysis were performed at the indicated time points. (C) Representative time-dependent phase-contrast images of AKTP organoids with the HUVEC layer treated with follistatin or its vehicle alone. Invadopodia are indicated by the arrowheads. Scale bars represent 100 μm. (D) Quantification of invadopodium formation on AKTP organoids under coculture with HUVECs treated without (control) or with follistatin ( n = 3 for biological chip replicates). (E) (i) Representative phase-contrast images (top, bars: 100 μm) and CLSM images (second to bottom, bars: 20 μm) of the immunostained HUVEC layer with AKTP organoids after six days of treatment without (control) or with follistatin. (ii) Quantification of SM22 intensity and endothelial layer disruption ( n = 3). (F) Relative mRNA levels of mesenchymal markers in HUVECs with AKTP organoids after six days of treatment without (control) or with follistatin ( n = 3 for biological chip replicates). (G) Relative mRNA levels of TGF-β family ligands in HUVECs after six days of treatment without (control) or with follistatin. Inhibin βA is a subunit of activin. ( n = 3 for biological chip replicates) The data in D, F and G are represented as mean ± S.D. Significant differences between control and treated condition were analyzed by two-way ANOVA in D or two-tailed unpaired Student’s t test in F and G, respectively. p values are provided. n.s.: not significant.

Article Snippet: For activin inhibition, we introduced 200 ng/mL follistatin (4889-FN-025, R&D Systems), an antagonist for activin ( 29 ), or 0.1% BSA in PBS as a vehicle.

Techniques: Inhibition, In Vitro, Control, Expressing, Disruption, Two Tailed Test