Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: The effects of ventricular pressure on AAC ‐induced atrial hypertrophy and atrial dilatation in ALK 4 +/− mice and WT littermates. A, Changes in ALK 4 protein expression by Western blotting in the left atria of ALK 4 +/− mice and WT littermates at 4 or 8 weeks post‐ AAC (n=5 in each group). B, LVEDP and (C) the left atrium weight‐to–tibia length ratio ( LA weight/ TL ) were compared between the 2 genotypes at 4 or 8 weeks post‐ AAC (n=8 in each group). D, Representative images of WGA staining of the left atrium tissues obtained at 4 or 8 weeks post‐ AAC in ALK 4 +/− mice and WT littermates. The scale bar is 20 μm. E, Quantification of the left atrium myocyte cross‐sectional area based on WGA staining (n=6 in each group, ≥100 cells per group) in ALK 4 +/− mice and WT littermates between sham and AAC groups. F, Left atrium length measured by echocardiography in indicated groups (n=8 in each group). G, mRNA expression of ANP , BNP , and GDF 15 of left atria from ALK 4 +/− mice and WT littermates after sham surgery or AAC (n=4 in each group). Data are presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05, ## P <0.01 vs corresponding WT group. AAC indicates abdominal aortic constriction; ALK4, Activin receptor‐like kinase 4; ANP, atrial natriuretic peptide; BNP, brain natriuretic peptide; GDF15, growth differentiation factor 15; LA, left atrium; LVEDP, left ventricle end‐diastolic pressure; NS, not significant; TL, tibia length; 4W, 4 weeks postprocedure; 8W, 8 weeks postprocedure; WGA, wheat germ agglutinin; WT, wild type.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: Expressing, Western Blot, Staining

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: Haplodeficient ALK 4 mitigated AAC ‐induced atrial fibrosis. A, Representative results of picrosirius red staining of the left atrium sections obtained from WT littermates and ALK 4 +/− mice at 4 or 8 weeks post‐ AAC (n=6 in each group; scale bar, 200 μm for all panels), and (B) statistical results of interstitial fibrosis area in left atrium (n=6 in each group; ≥40 fields per group). C, Relative mRNA level of collagen I in left atria obtained from ALK 4 +/− mice and WT littermates post‐ AAC (n=5 in each group). D, Changes in α‐ SMA and PCNA protein expression by Western blotting in the left atria of mice that underwent sham surgery or AAC in indicated groups (n=4 in each group). E, Representative images and quantitative results of staining Ki67 (green), α‐ SMA (red), and DAPI (blue) in left atrium sections of the indicated groups (n=6 in each group, ≥20 fields per group. Scale bar 50 μm). Data are presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05, ## P <0.01 vs corresponding WT group. AAC indicates abdominal aortic constriction; ALK4, Activin receptor‐like kinase 4; NS, not significant; PCNA, proliferation cellular nuclear antigen; α‐SMA, α‐smooth muscle actin; 4W, 4 weeks postprocedure; 8W, 8 weeks postprocedure; WT, wild type.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: Staining, Expressing, Western Blot

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: ALK 4 haplodeficiency blocked AAC ‐induced proliferation, differentiation, and extracellular matrix synthesis capacity of atrial fibroblasts. A, CCK ‐8 assay (Dojindo olecular Technologies, Rockville, MD) of atrial fibroblast extracted from the ALK 4 +/− mice post‐ AAC , WT littermates post‐ AAC , and their corresponding sham group at 0, 12, 24, and 48 hours (n=5 in each group). B, Changes in α‐ SMA and CTGF protein expression by Western blotting in the atrial fibroblasts extracted from mice that underwent sham operations or AAC in indicated groups (n=4 in each group). Data are presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05, ## P <0.01 vs corresponding WT group. AAC indicates abdominal aortic constriction; ALK4, Activin receptor‐like kinase 4; α‐ SMA , α‐smooth muscle actin; CTGF , connective tissue growth factor; NS, nonsignificant; 8W, 8 weeks postprocedure.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: CCK-8 Assay, Expressing, Western Blot

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: Haplodeficient ALK 4 protected against AAC ‐induced alterations of gap junctions in atrium. A, Changes of Cx40, Cx43 expression and (B) the ratio of Cx40 expression to Cx43 expression in left atrium obtained from ALK 4 +/− mice and WT littermates at 8 weeks post‐ AAC by Western blotting (n=5 in each group). C, Representative immunofluorescence images and quantitative data on Cx40 expression in left atrium sections of the indicated groups (n=4 in each group, ≥10 fields per group. Scale bar 50 μm). D, Representative immunohistochemistry image and quantitative results of Cx43 staining in left atrium sections obtained from ALK 4 +/− mice and WT littermates at 8 weeks post‐ AAC . The black arrows indicate that Cx43 is localized in dense clusters at the ends of the cells, and the red arrows indicate that distribution of Cx43 markedly changes to the lateral side of the cells. (n=4 in each group. Scale bar 20 μm). Data are presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05, ## P <0.01 vs corresponding WT group. AAC indicates abdominal aortic constriction; ALK4, Activin receptor‐like kinase 4; CTGF , connective tissue growth factor; Cx40, connexin 40; Cx43, connexin 43; NS , not significant; 8W, 8 weeks postprocedure; WT, wild type.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: Expressing, Western Blot, Immunofluorescence, Immunohistochemistry, Staining

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: ALK 4 haplodeficiency protected against AAC ‐induced inflammation in atria. A and B, Relative mRNA level of IL ‐6 and MCP ‐1 in left atria obtained from ALK 4 +/− mice and WT littermates post‐ AAC (n=4 in each group). C, Transwell assays were performed to determine AAC ‐induced chemotaxis of macrophages in each group. Atrial fibroblasts extracted from WT littermates post‐ AAC induced obvious chemotaxis of macrophages extracted from WT littermates post‐ AAC . However, atrial fibroblasts extracted from ALK 4 +/− mice post‐ AAC significantly blocked chemotaxis of macrophages extracted from ALK 4 +/− mice post‐ AAC (n=4 in each group. Scale bar 100 μm). Data are presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05, ## P <0.01 vs corresponding WT group. AAC indicates abdominal aortic constriction; ALK4, Activin receptor‐like kinase 4; Cx40, connexin 40; Cx43, connexin 43; IL ‐6, interleukin‐6; MCP ‐1, monocyte chemotactic protein 1; NS , not significant; 8W, 8 weeks postprocedure; WT, wild type.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: Chemotaxis Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: ALK 4 haplodeficiency protects against AAC ‐induced electrical remodeling. A, Expression level of ion channels related to electrical remodeling in atrium. Relative mRNA level of CACNA 1C, SCN 5A, KCND 2, KCND 3, KCNQ 1, KCNJ 2, KCNJ 3, KCNJ 12, and KCNA 5 in left atria obtained from ALK 4 +/− mice and WT littermates post‐ AAC (n=4 in each group). B, Changes of CACNA 1C and SCN 5A expression in left atria obtained from ALK 4 +/− mice and WT littermates at 8 weeks post‐ AAC by Western blotting (n=5 in each group). Data are presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05, ## P <0.01 vs corresponding WT group. AAC indicates abdominal aortic constriction; ALK4, Activin receptor‐like kinase 4; CACNA1C, calcium voltage‐gated channel subunit α1c; IL ‐6, interleukin‐6; KCNA5, potassium voltage‐gated channel, subfamily A, member 5; KCND2, potassium voltage‐gated channel, Shal‐related family, member 2; KCND3, potassium voltage‐gated channel, Shal‐related family, member 3; KCNJ12, potassium voltage‐gated channel, subfamily J, member 12; KCNJ2, potassium voltage‐gated channel, subfamily J, member 2; KCNJ3, potassium voltage‐gated channel, subfamily J, member 3; KCNQ1, potassium voltage‐gated channel, subfamily Q, member 1; MCP ‐1, monocyte chemotactic protein 1; NS, not significant; SCN5A, sodium voltage‐gated channel α subunit 5; 8W, 8 weeks postprocedure; WT, wild type.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: Expressing, Western Blot

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: ALK 4 haplodeficiency inhibited AAC ‐induced decreasd I N a , I C a‐L , I K 1 current densities and the accompanying APD shortening in atrial myocytes. A, Whole‐cell membrane capacitance of atrial myocytes isolated from ALK 4 +/− mice and WT littermates that underwent sham surgery or AAC (n=10 in each group). B, AP s recorded from a representative atrial myocyte in each group at a pacing rate of 1 Hz. C, Averaged data of APD at 90% repolarization ( APD 90 ) and APD at 50% repolarization ( APD 50 ) under referred conditions. D, Representative original traces of whole‐cell I to current (300 ms depolarization step pulses from −40 to +50 mV with a step size of +10 mV) and representative density‐voltage curve for I to in atrial myocyte isolated from mice in each group. Insert, voltage protocol (n=8 in each group). E, Representative recordings of whole‐cell I N a current (sweep 1‐sweep 16) in atrial myocyte isolated from mice in each group. Insert, voltage protocol. The lower panel shows representative I N a density‐voltage relationships in each group (n=8 in each group). F, Representative recordings of whole‐cell I C a‐L current in atrial myocyte isolated from mice in each group. Insert, voltage protocol. The lower panel shows representative I C a‐L current density‐voltage relationships in each group (n=8 in each group). G, Original representative recordings of the inwardly rectifier I K 1 current (only voltage steps between −120 and −40 mV with a step size of +10 mV were shown) and representative density‐voltage curve for I K 1 in atrial myocyte isolated from mice in each group. Insert, voltage protocol. (n=8 in each group). Data were presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05, ## P <0.01 vs corresponding WT group. ALK4 indicates Activin receptor‐like kinase 4; AP , action potential; NS, not significant; 8W, 8 weeks postprocedure.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: Isolation

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: AF inducibility and duration in ALK 4 +/− mice and WT littermates subjected to sham or AAC operation. A, Heart rate and (B) P wave duration measured in vivo electrophysiological investigations using ALK 4 +/− mice and WT littermates after sham operation or AAC (n=6 in each group). C and D, Example electrical tracings of surface and intracardiac leads from (C) WT littermates and (D) ALK 4 +/− mice post‐ AAC during electrophysiological studies with cycle length of 40 milliseconds. Scale 100 milliseconds. E, Number and (F) total duration of AF episodes during the electrophysiological studies (n=15 in each group). Data are presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05 vs corresponding WT group. AAC indicates abdominal aortic constriction; AF, atrial fibrillation; ALK4, Activin receptor‐like kinase 4; HR, heart rate; NS, not significant; SR, sinus rhythm; 8W, 8 weeks postprocedure; WT, wild type.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: In Vivo

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: ALK 4 haplodeficiency significantly blocks activation of Smad2/3 pathway in atrium. A, Representative Western blotting and quantitative data of (B) phosphorylation protein levels of Smad2, Smad3, (C) ERK , JNK and P38 of left atria from ALK 4 +/− mice and WT littermates after sham surgery or AAC (n=4 in each group). Data are presented as mean± SEM . * P <0.05 vs corresponding sham group. # P <0.05 vs corresponding WT group. AAC indicates abdominal aortic constriction; NS , not significant; ALK4, Activin receptor‐like kinase 4; 8W, 8 weeks postsurgery; WT, wild type.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: Activation Assay, Western Blot

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Activin Receptor‐Like Kinase 4 Haplodeficiency Mitigates Arrhythmogenic Atrial Remodeling and Vulnerability to Atrial Fibrillation in Cardiac Pathological Hypertrophy

doi: 10.1161/JAHA.118.008842

Figure Lengend Snippet: Intraperitoneal administration of ALK 4 inhibitor SB ‐431542 inhibited atrial interstitial fibrosis, improved the expression of functional Cx40 protein and CACNA 1C mRNA in atrium, and inhibited the vulnerability to AF at 8 weeks post‐ AAC . A, Representative results of picrosirius red staining of the left atrium sections obtained from WT littermates subjected to sham surgery or AAC or AAC operation with intraperitoneally administrated with SB ‐431542 (1 μmol/L solution, 100 μL/animal) or vehicle alone on first and third day and every 4 days after the production of AAC for 8 weeks (n=15 in each group; scale bar 200 μm). B, Representative western blots and quantitative data of Cx40 in left atrium obtained at 8 weeks post AAC in indicated groups (n=4 in each group). C, Statistical results of interstitial fibrosis area in left atrium (n=5 in each group). D, mRNA expression of CACNA 1C in left atria from referred groups (n=4 in each group). E and F, Example electrical tracings of surface and intracardiac leads from (C) WT littermates post AAC and (D) WT littermates post AAC with intraperitoneal administrated with SB ‐431542 during electrophysiological studies with cycle length of 30 ms. Scale, 100 ms. G, Number of AF ‐episodes during the electrophysiological studies (n=15 in each group). Data are presented as mean± SEM . * P <0.05, ** P <0.01 vs corresponding sham group. # P <0.05 vs corresponding AAC group. † P <0.05 vs corresponding AAC plus vehicle group. AAC indicates abdominal aortic constriction; ALK4, Activin receptor‐like kinase 4; CACNA1C, calcium voltage‐gated channel subunit α1c; Cx40, connexin 40; NS, not significant; SR, sinus rhythm; 8W, 8 weeks postsurgery; WT, wild type.

Article Snippet: After the animal surgery, ALK4 inhibitor SB‐431542 (1 μmol/L solution, 100 μL/mouse; Selleckchem, Houston, TX) or vehicle alone was intraperitoneally administered on the first and third days after the models were established successfully and was repeated every 4 days for a further 8 weeks.,

Techniques: Expressing, Functional Assay, Staining, Western Blot

Journal: International journal of biological sciences

Article Title: Beauvericin suppresses the proliferation and pulmonary metastasis of osteosarcoma by selectively inhibiting TGFBR2 pathway.

doi: 10.7150/ijbs.86214

Figure Lengend Snippet: Figure 4. BEA interacts with TGFBR2 and inhibits the TGFBR2-mediated aggressive progression of OS cells in vitro. (A) The binding of BEA to TGFBR2 and TFGBR1 was determined by DARTS approach. SB431542 was used as the negative control for TGFBR2 and the positive control for TGFBR1. (B) CETSA was used to evaluate

Article Snippet: SB431542 were obtained from TargetMol (T1726, Boston, MA, USA).

Techniques: In Vitro, Binding Assay, Negative Control, Positive Control