Rabbit anti pdcd4

H9C2 cells were cultured in DMEM/F12 (Hyclone, Logan, UT, USA) medium supplemented with 10% FBS, H9C2 cells were pre-incubated with 10% Exo-FBS DMEM with stressed, normal or without CPC-exosomes (2 × 10⁹ particles per ml) for 24 h, then treated with 100 μM H₂O₂ for 6 h. Following treatment, the apoptosis rate analyzed by the Flow cytometry with Annexin V/PI kit (BD Bioscience, Franklin Lakes, NJ, USA), according to the instructions of the manufacturer. Whole-cell lysate were prepared by adding cell lysis buffer (Thermo), cell protein concentration determined by BCA Protein Assay Kit (Thermo), and then resolved on a 10% sodium dodecyl sulfate bis-tris gel, and transferred to an Immobilon FL PVDF membrane (Millipore). The membrane was blocked with 5% non-fat milk in TBST buffer, and incubated with rabbit anti-caspase-3 (1 : 1000, #9662, Cell Signaling Technology), rabbit anti-PDCD4(1 : 1000, #9535, Cell Signaling Technology) overnight, and then, incubated with HRP linked goat anti-rabbit IgG (1 : 5000, Cell Signaling Technology), and the protein bands were visualized using the with automatic imager (General Electric). The blots were quantified using FluorChem 8900 software (Alpha Innotech Corporation, San Leandro, CA, USA), and the relative protein expression was normalized to β-actin.