Journal: Cardiovascular Research

Article Title: Ecstasy produces left ventricular dysfunction and oxidative stress in rats

doi: 10.1093/cvr/cvn129

Figure Lengend Snippet: 3,4-methylenedioxymethamphetamine increases tyrosine nitration of SERCA-2a. (A) Nitrotyrosine immunoprecipitates from saline (n = 5 pooled) and 3,4-methylenedioxymethamphetamine (n = 6 pooled) left ventricular lysates were probed by western blot analysis using SERCA-2a antibody. (B) SERCA-2a protein levels normalized to GAPDH (n = 5). (C) SERCA-2a protein levels normalized to phospholamban (n = 5).

Article Snippet: Immunoprecipitated proteins were subjected to western blot analysis with SERCA-2a (Biomol) antibodies.

Techniques: Nitration, Western Blot

Journal: BMC Cardiovascular Disorders

Article Title: Exercise training restores the cardiac microRNA-1 and −214 levels regulating Ca 2+ handling after myocardial infarction

doi: 10.1186/s12872-015-0156-4

Figure Lengend Snippet: Expression of cardiacmRNA of the Serca-2a ( a ), mRNA of the NCX ( b ), miRNA-1 ( c ) and −214 ( d ) by real-time PCR reaction and expression of NCX ( e ) and Serca-2a protein expression ( f ) by western blot (blot above showed), in remote myocardium of the infarcted. Percentage related to SED SHAM group, n = 7 each group. All results are presented as mean ± standard deviation. Different letters indicate statistically different groups ( P < 0.05)

Article Snippet: The blot membrane was then incubated in a blocking buffer for 2 h at room temperature and then incubated overnight at 4 °C with Serca-2a (catalog No. #4388) monoclonal antibody (Cell Signaling Tech., MA, USA) or total PLP (catalog No. ab86930) polyclonal antibody or corresponding phosphorylated PLP (Ser 16 ) (catalog No. ab15000) polyclonal antibody or NCX (catalog No. ab2869) monoclonal antibody or GAPDH (catalog No.ab9484) monoclonal antibody (Abcam, Cambridge, UK).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Standard Deviation

Journal: BMC Cardiovascular Disorders

Article Title: Exercise training restores the cardiac microRNA-1 and −214 levels regulating Ca 2+ handling after myocardial infarction

doi: 10.1186/s12872-015-0156-4

Figure Lengend Snippet: Correlation between ( a ) miRNA-214 levels and Serca-2a protein expressionby western blot and ( b ) VO 2 max and miRNA-214 levels. Blue lines show linear regression applied for SHAM rats and red lines for INF rats, n = 7 each group

Article Snippet: The blot membrane was then incubated in a blocking buffer for 2 h at room temperature and then incubated overnight at 4 °C with Serca-2a (catalog No. #4388) monoclonal antibody (Cell Signaling Tech., MA, USA) or total PLP (catalog No. ab86930) polyclonal antibody or corresponding phosphorylated PLP (Ser 16 ) (catalog No. ab15000) polyclonal antibody or NCX (catalog No. ab2869) monoclonal antibody or GAPDH (catalog No.ab9484) monoclonal antibody (Abcam, Cambridge, UK).

Techniques: Western Blot

Journal: BMC Cardiovascular Disorders

Article Title: Exercise training restores the cardiac microRNA-1 and −214 levels regulating Ca 2+ handling after myocardial infarction

doi: 10.1186/s12872-015-0156-4

Figure Lengend Snippet: Expression of cardiac phospholamban protein expression ( a ) by western blot (blot above showed) and Serca-2a activity ( b ) by measurement of Ca 2+ -ATPase activity in remote myocardium of the infarcted. Percentage related to SED SHAM group, n = 7 each group. All results are presented as mean ± standard deviation. Different letters indicate statistically different groups ( P < 0.05)

Article Snippet: The blot membrane was then incubated in a blocking buffer for 2 h at room temperature and then incubated overnight at 4 °C with Serca-2a (catalog No. #4388) monoclonal antibody (Cell Signaling Tech., MA, USA) or total PLP (catalog No. ab86930) polyclonal antibody or corresponding phosphorylated PLP (Ser 16 ) (catalog No. ab15000) polyclonal antibody or NCX (catalog No. ab2869) monoclonal antibody or GAPDH (catalog No.ab9484) monoclonal antibody (Abcam, Cambridge, UK).

Techniques: Expressing, Western Blot, Activity Assay, Standard Deviation

Journal: Cardiovascular Diagnosis and Therapy

Article Title: In vivo effects of nitrosyl hydrogen on cardiac function and sarcoplasmic reticulum calcium pump (SERCA2a) in rats with heart failure after myocardial infarction

doi: 10.21037/cdt-20-201

Figure Lengend Snippet: Effects of nitroxyl hydrogen on protein expression of SERCA2a. Values are means ± SD, *P<0.01 vs. sham. ΔP<0.05 vs. HF.

Article Snippet: Main experimental reagents Angeli’s salt (Cayman), rat amino-terminal forebrain natriuretic peptide ELISA kit (Abnova), cell lysate, protease inhibitor, BCA protein concentration assay kit (all from Wuhan Boster), SERCA2a primary antibody (Abnova), goat anti-mouse secondary antibody (Abnova), β-actin primary antibody (Abnova), nitrocellulose membrane (Abnova), ATPase assay kit (Nanjing Jiancheng Institute of Bioengineering).

Techniques: Expressing

Journal: Cardiovascular research

Article Title: Chronic depletion of glutathione exacerbates ventricular remodelling and dysfunction in the pressure-overloaded heart.

doi: 10.1093/cvr/cvs333

Figure Lengend Snippet: Figure 3 Ca2+ uptake and SERCA2a expression and glutathionylation in myocardium and cell shortening and [Ca2+]i transient in cultured mouse cardiomyocytes. (A) SERCA-specific Ca2+ uptake in the supernatant of myocardial homogenates. (B) SERCA2a mRNA expression in the myocardium, normalized to GAPDH mRNA expression. (C) Glutathionylation levels of SERCA2a in the myocardium. The band intensity of the immunoblotting using anti-GSH antibody was expressed as a ratio of that using anti-SERCA2a antibody and normalized to the sham-operated GCLM+/+ myocardium (¼1). Upper insets show representative immunoblots of the immunoprecipitated SERCA2a from myocardial homogenates using anti-GSH antibody or anti-SERCA2a antibody under reduced or non-reduced conditions. (D) Cell shortening of cultured cardiomyocytes in response to field electrical stimulation. Peak (E) and time to 50% decay (F) of [Ca2+]i transients of cultured cardiomyocytes in response to electrical stimulation. n ¼ 6–8 in each experiment. †P , 0.05 compared with the respective sham-operated group. *P , 0.05.

Article Snippet: For detection of glutathionylated SERCA2a using immunoblot analysis, myocardial tissue homogenates were immunoprecipitated by a cross-link immunoprecipitation kit (Thermo Fisher Scientific, San Jose, CA, USA) using rabbit polyclonal SERCA2a antibody (#4219S, Cell signaling technology, Beverly, MA, USA).

Techniques: Expressing, Cell Culture, Western Blot, Immunoprecipitation

Journal: Cardiovascular research

Article Title: Chronic depletion of glutathione exacerbates ventricular remodelling and dysfunction in the pressure-overloaded heart.

doi: 10.1093/cvr/cvs333

Figure Lengend Snippet: Figure 6 Effects of GSH-EE supplementation in TAC experiments. (A–G) LV end-diastolic and end-systolic dimensions (LVDd and LVDs), fractional shortening (%LVFS) on echocardiography, myocardial fibrotic area and microvessel density, and glutathionylation levels of SERCA2a, and SERCA-specific Ca2+ uptake, respectively, in GCLM2/2 mice after treatment with GSH-EE or PBS as a placebo and in GCLM+/+ mice with no treat- ment during the 4 weeks of TAC. (H and I) GSH content and GSSG content in myocardium, respectively, after treatment with intraperitoneal injection of GSH-EE or PBS for the last week of 4 weeks of TAC. 2/2 denotes GCLM2/2 mice, and +/+ denotes GCLM+/+ mice. n ¼ 6 in each experiment. *P , 0.05.

Article Snippet: For detection of glutathionylated SERCA2a using immunoblot analysis, myocardial tissue homogenates were immunoprecipitated by a cross-link immunoprecipitation kit (Thermo Fisher Scientific, San Jose, CA, USA) using rabbit polyclonal SERCA2a antibody (#4219S, Cell signaling technology, Beverly, MA, USA).

Techniques: Injection