Quantitative Analysis of Cardiac Signaling Pathways

Total protein and nuclear fraction were extracted from cardiomyocytes and heart tissue and Western blotting was performed as previously described [52 (link),55 (link)]. Ten µg of each fraction were resolved by SDS-PAGE. For Western blotting, anti-phospho-ERK1/2 (T202/Y204) antibody (Cell Signaling Technology, Danvers, MA, USA), anti-ERK1/2 antibody (Cell Signaling Technology), anti-phospho-GATA4 (S105) antibody (Abcam, Cambridge, United Kingdom), and anti-GATA4 antibody (Cell Signaling Technology) were used as primary antibodies, and anti-mouse antibody (MBL, Aichi, Japan) and anti-rabbit antibody (MBL) were used as secondary antibodies. Western blotting signals were visualized with an Amersham Imager 680 blot imager (GE Healthcare, Chicago, IL, USA) and quantified with NHI ImageJ software (version 1.52a).

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Shimizu K., Sunagawa Y., Funamoto M., Honda H., Katanasaka Y., Murai N., Kawase Y., Hirako Y., Katagiri T., Yabe H., Shimizu S., Sari N., Wada H., Hasegawa K, & Morimoto T. (2021). The Selective Serotonin 2A Receptor Antagonist Sarpogrelate Prevents Cardiac Hypertrophy and Systolic Dysfunction via Inhibition of the ERK1/2–GATA4 Signaling Pathway. Pharmaceuticals, 14(12), 1268.

Publication 2021

anti-phospho-ERK1/2 (T202/Y204) antibody anti-ERK1/2 antibody

Anti antibody Antibodies Antibody Cardiomyocytes Erk1 Heart Mouse Protein Rabbit Sds page Tissue

Corresponding Organization : Shizuoka General Hospital

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Variable analysis

independent variables

Total protein and nuclear fraction extracted from cardiomyocytes and heart tissue

dependent variables

Phosphorylation levels of ERK1/2 (T202/Y204)
Phosphorylation levels of GATA4 (S105)

control variables

10 µg of each fraction resolved by SDS-PAGE
Anti-phospho-ERK1/2 (T202/Y204) antibody, anti-ERK1/2 antibody, anti-phospho-GATA4 (S105) antibody, and anti-GATA4 antibody used as primary antibodies
Anti-mouse antibody and anti-rabbit antibody used as secondary antibodies
Western blotting signals visualized with Amersham Imager 680 blot imager and quantified with NHI ImageJ software (version 1.52a)

positive controls

None specified

negative controls

None specified

Annotations

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