Quantifying Heart-Related Gene Expression

Using the Total RNA Isolation Reagent (BS259A, Biosharp, China), total RNA was extracted from heart tissues or cells. After that, the total RNA underwent reverse transcription and quantification on a Real-Time PCR system (7500, ThermoFisher, USA) using the One-Step SYBR Green qRT-PCR Kit (BL643B, Biosharp, China). Under the adoption of the 2^−ΔΔCT method, B-type natriuretic peptide (BNP), atrial natriuretic peptide (ANP), major histocompatibility complex-B (B-MHC), GSK3A, CD36, PDK4 (Pyruvate dehydrogenase kinase 4), and Cpt1b (carnitine palmitoyltransferase 1B) expressions were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) [12 (link)]. Supplementary Table 3 lists the primer sequences.