Extended Data Fig. 5: JOSD2 regulates the ubiquitination of SERCA2a.
(a) Myc-SERCA2a and HA-Ub were transfected into cardiomyocytes together with Flag-JOSD2 and then subjected to 10 μM MG132 for 2 h. Lysates were subjected to Co-IP with anti-Myc, which was followed by immunoblot with an anti-HA (n = 3). (b) Myc-SERCA2a and HA-Ub were transfected into cardiomyocytes together with siJOSD2 and then subjected to 10 μM MG132 for 2 h. Lysates were subjected to Co-IP with anti-Myc, which was followed by immunoblot with an anti-HA (n = 3). (c) Lysates of heart tissue from Josd2−/− mice were subjected to Co-IP with anti-SERCA2a, which was followed by immunoblot with an anti-Ub (n = 3). (d) Myc-SERCA2a and HA-K63 were transfected into NIH/3T3 together with Flag-JOSD2 and then subjected to 10 μM MG132 for 2 h. Lysates were subjected to Co-IP with anti-Myc, which was followed by immunoblot with an anti-HA (n = 3). (e) General E3 enzymes ubiquitination prediction for SERCA2a (GPSUber: http://gpsuber.biocuckoo.cn/index.php).
