Ab110056

Western blot was performed as previously described (29 (link)). Briefly, sample lysates were resolved in SDS-PAGE and transferred onto PVDF membrane (Merck Millipore, Billerica, MA, USA). The PVDF membranes were incubated with the primary antibody JP2 (ab110056, Abcam, MA, USA, 1:1000) and GAPDH (HA-ET1702-66-200, HUABIO, Hangzhou, China), respectively. Then samples were incubated with HRP-conjugated secondary antibodies (LK-GAM007, LK-GAR007, LK-RAG007, MULTISCIENCES, Shanghai, China). Blots were developed using enhanced chemiluminescence reagents (Cat#. 32106, Pierce™, IL, USA).

All immunostaining was conducted as previously described (29 (link)). Briefly, samples were probed with the primary antibodies as follows: cardiac troponin T (cTnT, ab8295, Abcam, MA, USA.1:100), JP2 and its N-terminal fragment (NT-JP2) (ab110056, Abcam, MA, USA, 1:100). Then samples were incubated with DyLight488 goat anti-mouse secondary antibody (GAM4882, Multisciences, (Lianke) Biotech Co., Ltd. Hangzhou, China.1:200) and 4′, 6-Diamidino-2-phenylindoledihy-drochloride (DAPI, ab188804, Abcam, MA, USA. 2 μg/ml). Images were acquired under laser scanning Confocal microscopes from Leica Microsystems (Leica Microsystems Inc., Buffalo Grove, IL, USA). Relative ratio of JP2-NT nuclear translocation was analyzed by ImageJ software (National Institutes of Health, USA).