2.6. Western blot analysis

The proteins from heart tissues were divided. Primary antibodies used included p44/42 MAPK (Erk1/2) (1:1000; 4695, Cell Signaling Technology, USA), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (1:1000; 4370, Cell Signaling Technology, USA), SAPK/JNK (1:1000; 9252, Cell Signaling Technology, USA), Phospho-SAPK/JNK (Thr183/Tyr185) (1:1000; 4668, Cell Signaling Technology, USA), p38 MAPK (1:1000; 8690, Cell Signaling Technology, USA), Phospho-p38 MAPK (Thr180/Tyr182) (1:1000; 4511, Cell Signaling Technology, USA), Smad2 (1:500; 5339, Cell Signaling Technology, USA), Phospho-Smad2 (Ser465/Ser467) (1:500; 18338, Cell Signaling Technology, USA), Smad2/3 (1:1000; 8685, Cell Signaling Technology, USA), Phospho-Smad2 (Ser465/467)/Smad3 (1:1000; Ser423/425) (8828, Cell Signaling Technology, USA), TGF-β (1:500; 3709, Cell Signaling Technology, USA), and GAPDH (1:1000; 2118, Cell Signaling Technology, USA). The secondary antibody used for immunoblotting was Anti-rabbit IgG, HRP-linked Antibody (1:3000; 7074, Cell Signaling Technology, USA).