Extended Data Fig. 5: NMT5 S-nitrosylates ACE2 in vitro and in vivo.
From: Targeted protein S-nitrosylation of ACE2 inhibits SARS-CoV-2 infection
a, Detection of SNO-ACE2 in vitro. HeLa-ACE2 cells were exposed to 10 μM NMT3 or 5 μM NMT5. After 1 h, cells were subjected to the biotin-switch assay in the presence or absence of ascorbate. SNO-ACE2 and input ACE2 were detected by immunoblotting with anti-ACE2 antibody. b, c, Ratio of SNO-ACE2/input ACE2. Data are mean + s.e.m. by two-tailed Student’s t test. n = 5 biological replicates. d–i, Detection of SNO-ACE2 in vivo. Syrian hamsters received 10 mg/kg of NMT3 or of NMT5 by oral gavage and were sacrificed 48 h later. Kidney and lung tissues were subjected to biotin-switch assay in the presence or absence of ascorbate. Note that in some samples, low levels of SNO-ACE2 were observed in control tissue, suggesting endogenous S-nitrosylation of ACE2 may occur at low levels. Graphs show ratio of SNO-ACE2/input ACE2. Data are mean + s.e.m. by two-tailed Student’s t test. n = 3 Syrian hamsters for each condition.
