2.7 Western blotting assay
Western blotting was conducted using material extracted from the atrium, as previously described (Feng et al., 2020; Xu et al., 2012; Xu et al., 2010). Briefly, atrial tissue was homogenized with PRO-PREP protein extract (iNtRON Biotechnology Inc., Kyungki-Do, Korea), and 10 µg of protein was subjected to SDS-PAGE on a polyacrylamide gel (Bio-Rad Laboratory, Hercules, CA, USA). Subsequently, the proteins were transferred to the membrane using semi-dry electroblotting. The resulting membranes were incubated overnight at 4°C with the following primary antibodies: p-RyR2(Ser2814) (A010-31AP; Badrilla), ox-CaMKII (07-1387; Merck), CACNA1C (ab84814; Abcam), TRPC3 (ab51560; Abcam), TRPC6 (ab62461; Abcam), p-JNK (#4671S; Cell Signaling Technology), JNK (#9258; Cell Signaling Technology), p-ERK (#9101S; Cell Signaling Technology), ERK (#9102; Cell Signaling Technology), p-NF-κB p65 (Ser536) (#3033; Cell Signaling Technology), and GAPDH (#2118S; Cell Signaling Technology). The membranes were then incubated with the appropriate secondary antibodies, namely HRP-conjugated goat anti-rabbit IgG (ab6721; Abcam) or HRP-conjugated rabbit anti-mouse IgG (ab97046; Abcam), for 1 h at room temperature. Immunoreactions were detected using enhanced chemiluminescence (ECL Prime western blotting Detection; GE Healthcare).