Anti pkcγ

The following primary antibodies were used in this study: anti-NR1 (#MAB1586 Millipore), anti-NR1C1 (#AB5046P Millipore), anti-NR2A (#ab14596 Abcam), anti-NR2B (#ab14400 Abcam), anti-NR1 C1 P-S897 (#ABN99 Millipore), anti-NR2B P-Y1472 (#AB5403 Millipore), anti-NR2B P-Y1303 (#ab81271 Abcam), anti-NR2B P-Y1480 (#PA1-4733 ThermoFisher Scientific), anti NR2B P-Y1252 (#ab18532 Abcam), anti-GSK3β (#9315 Cell Signaling Technology), anti-P-S9 GSK3β (#9336 Cell Signaling Technology), anti-P-Y216 GSK3β (#ab75745 Abcam), anti-Akt (#4691 Cell Signaling Technology), anti-P-S473 Akt (#4060 Cell Signaling Technology), anti-P-T308 Akt (#2965 Cell Signaling Technology), anti-PKC sampler kit (#611421 BD Biosciences), anti-PKCγ (#ab71558 Abcam), and anti-α tubulin (#T9026 Sigma Aldrich).

Both H9C2 cells and myocardial tissues were harvested for Western blot analysis (Western Blot Detection Kit, Elabscience) following standard protocol according to the manufacturer’s instructions and Towbin system buffer was used. Primary antibodies used in Western blot were as follows: anti-GLP-1R (1:200, Abcam, UK), anti-Akt (1:200, Abcam, UK), anti-p-Akt (1:200, Abcam, UK), anti-PI3K (1:500, Santa Cruz, USA), anti-PKC (1:500, Abcam, UK), anti-PKCα (1:500, Abcam, UK), anti-PKCβ (1:500, Abcam, UK), anti-PKCγ (1:500, Abcam, UK), anti-PKCδ (1:500, Abcam, UK) and anti-β-actin (1:500, Santa Cruz, USA). Secondary antibodies were horseradish peroxidase-conjugated goat anti-rabbit lgG (Santa Cruz, USA) at 1:5000 dilution. The results were visualized using an enhanced chemiluminescence system (ECL, Amersham). Densitometric analysis was conducted using ImageJ software (NIH, USA).