2.9. Western‐blot analysis

The protein samples were prepared from the cells and tissues as previously described. 20 Equal amount of cardiac cell lysates were run on SDS‐polyacrylamide gels and incubated with antibodies against proteins Mfn1 (sc‐166644, 1:1000), Mfn2 (sc‐100560, 1:1000), Drp1 (sc‐271583, 1:500), SERCA2 (SC‐376235, 1:500), NCX (Santa Cruz, sc‐12972), phospho‐phospholamban‐Ser¹⁶ (pPLB; sc‐17024, 1:10 000), phospholamban (PLB; sc30142, 1:1000), SGLT2 (ab37296,1:1000). β‐actin (Santa Cruz, sc‐47778, 1:5000) and GAPDH (Cell Signaling, D16H11, 1/5000) were used as housekeeping proteins to detect equal protein loading. Immunoreactive bands were detected by a chemiluminescent reaction (ECL kit, Amersham Pharmacia, USA). The densities of the bands are analysed using ImageJ software, and the results were presented as fold changes.