NBR1 Antibody - #DF12049

Figure 4 ｜ Effect of bradykinin postconditioning on the immunopositivity of AMPK/ mTOR signaling pathway, autophagy-related proteins, and brain injury marker in rats with restoration of spontaneous circulation, as detected by immunohistochemistry.

Fig 6 MARCH6 recruits TOLLIP as an autophagy cargo receptor for NS5 degradation. (A and B) Lysates of DEFs co-transfected with the indicated Flag-tagged cargo receptors and HA-MARCH6 (A) or HA-NS5 (B) were subjected to co-IP assays with an anti-Flag antibody, followed by Western blotting analysis. (C–E) Lysates from DEFs co-transfected with plasmids encoding HA-NS5 and Flag-TOLLIP (C), Flag-OPTN (D), or Flag-NBR1 (E), together with or without MYC-MARCH6, and treated with Baf A1 (10 µM), were subjected to co-IP assays using an anti-Flag antibody, followed by immunoblot analysis with the indicated antibodies. (F–H) Lysates from DEFs co-transfected with specific siRNA targeting MARCH6 and plasmids encoding Flag-NS5 and HA-TOLLIP (F), HA-OPTN (G), or HA-NBR1 (H) were subjected to co-IP assays using an anti-Flag antibody, followed by immunoblot analysis with the indicated antibodies. (I–K) Immunoblot analysis of lysates of DEFs co-transfected with HA-NS5 and shTOLLIP (I), shOPTN (J), or shNBR1 (K), together with or without Flag-MARCH6 for 24 h. (L and M) DEFs were transfected with shNegative or shTOLLIP, along with Flag-MARCH6 or an empty vector for 24 h and then infected with TMUV at an MOI of 0.1. E protein expression was analyzed by Western blotting (L), and viral titers were quantified using the TCID50 assay (M). (M) Data are presented as mean values ± standard error of the mean of three independent experiments (Student’s t-test; **, P < 0.01) ns, not significant.

Figure 4 ｜ Effect of bradykinin postconditioning on the immunopositivity of AMPK/ mTOR signaling pathway, autophagy-related proteins, and brain injury marker in rats with restoration of spontaneous circulation, as detected by immunohistochemistry. Compared with the Sham group, p-mTOR and p62 immunopositivity in the ROSC group was significantly decreased, S100β, p-AMPK, and LC3 immunopositivity was significantly increased, and NBR1 immunopositivity remained unchanged. Compared with the ROSC group, p-mTOR, p62, and S100β immunopositivity in the BK group was significantly decreased, while p-AMPK, NBR1, and LC3 immunopositivity was significantly increased. Compared with the BK group, LC3, p-AMPK, and NBR1 immunopositivity in the CP+BK group was significantly decreased, whereas S100β, p62, and p-mTOR immunopositivity was significantly increased; LC3, p-AMPK, and NBR1 immunopositivity in the Ra+BK group was significantly increased, while S100β, p62, and p-mTOR immunopositivity was significantly decreased. Scale bars: 50 μm. All data are presented as the mean ± SD (n = 8). *P < 0.05, **P < 0.01, vs. Sham group; #P < 0.05, ##P < 0.01, vs. ROSC group; †P < 0.05, ††P < 0.01, vs. BK group (one-way analysis of variance followed by Tukey’s post hoc test). BK: Bradykinin; CP: compound C; IOD: integrated optical density; LC3: microtubule-associated protein 1 light chain 3; NBR1: neighbor of breast cancer 1 gene; p-AMPK: phosphorylated adenosine-monophosphate activated protein kinase; p-mTOR: p-mechanistic target of rapamycin; Ra: rapamycin; ROSC: restoration of spontaneous circulation; S100β: S100 calcium-binding protein B; Sham: sham operation.