Dulbeccos s modified eagle s medium dmem

Human bronchial epithelial cells (16HBE) and human embryonic kidney cells (293T) were obtained from the cell bank of the Chinese Academy of Sciences (Shanghai, China). 16HBE cells were cultured with RPMI 1640 media (Gibco, New York, USA) and 293T cells cultured with Dulbeccos’s modified Eagle’s medium (DMEM; Gibco), both containing 10% foetal bovine serum (FBS; Evergreen, Hangzhou, China), 100 mg/L penicillin, and 100 mg/L streptomycin (Gibco) in a 5% CO₂ incubator at 37°C. When cells grew to 70%-80% confluence, they were passaged using trypsin-ethylenediaminetetraacetic acid (Gibco).

Cardiomyocyte primary culture extracting kit was obtained from Worthington Biochemical Corporation (Lakewood, USA). Dulbeccos’s Modified Eagle’s Medium (DMEM), penicillin, streptomycin, amphotericin and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, USA). Cytosine β-D-arabinofuranoside (ARA-C), fibronectin, KN-93 (Ca²⁺/calmodulin-dependent protein kinase II inhibitor) and ultra-pure succinic acid were obtained from Sigma (St. Louis, USA), Fluo-4/AM, DAPI and secondary antibodies conjugated to Alexa-488, Alexa-633 and Silencer kit were from Ambion, Life Technologies (New York, USA). Trizol reagent was obtained from Invitrogen (Eugene, USA). Polyclonal anti- GAPDH, anti- HDAC5, and anti-ANP antibodies were from Santa Cruz Biotechnology (Santa Cruz, USA). Monoclonal anti – Phospho-p44/42 MAPK and polyclonal anti-MEK1/2 were purchased from Cell Signaling Technology (Boston, USA), monoclonal anti-CAMK2δ from Abnova (Taipei, Taiwan). Hydromount was from National Diagnostics (St. Louis, USA). PD- 98059 (MEK 1/2 innhibitor) was from Calbiochem (Germany). Enhanced chemiluminescence (ECL-plus Western Blotting Detection System) and peroxidase-conjugated antibodies were purchased from Amersham Biosciences (Buckinghamshire, UK). All other reagents were of the highest quality commercially available.