Fig. 7.
Increased ubiquitin E3 ligase activity after S-nitrosylation of RNF11, RNF41 and RNF141. A, In vitro ubiquitination activity assay with or without GSNO treatment for 1 h in presence of ubiquitin, E1 and E2 (UbcH5c) enzymes analyzed by immunoblot with anti-ubiquitin and anti-GST antibodies. B, In vivo ubiquitination activity assay with or without GSNO treatment for 6 h analyzed by immunoblot with anti-HA and anti-V5 antibodies. Actin and HA-ubiquitin expression were monitored as loading controls. All experiments were repeated three times.