Fig. 8

Inhibition of IKKβ abolished the effectiveness of IKKγ translocation. A, B The influence of C25-140 and TRAF6 the inhibition on IKK/IκBα/NF-κB proteins was detected when TRIM37 was overexpressed after 1 h to 36 h of reoxygenation. TRAF6 inhibition prematurely ended the 36-h spike in inflammation, but C25-140 did not. C Immunofluorescence analysis of IKKγ and F4/80 in primary Kupffer cells after H/R treatment and 24 h and 36 h of reoxygenation, scale bar = 50 μm for immunofluorescence. D The specific IKKβ inhibitor IMD0354 was used for further study. The influence of IMD0354 on IKK/IκBα/NF-κB protein was detected. E The underlying mechanism by which TRIM37 aggravates liver I/R injury through TRAF6 ubiquitination and IKKγ translocation. The activation of IKK inflammatory signaling pathway was induced by the activation of K63 ubiquitination derived from the binding of TRIM37 to TRAF6. Cytoplasmic translocation of IKKγ might be one of the main functional IKK subunits in this process. Subsequently, canonical NF-κB p65 is phosphorylated and transported into the nucleus, resulting in the release of inflammatory cytokines and chemokines. TRIM37: tripartite motif containing 37. TRAF6: tumor necrosis factor receptor-associated factor 6. IKK: inhibitor of nuclear factor kappa-B kinase. IκBα: inhibitor of NF-κB. NF-κB: nuclear factor kappa B. CXCL-10: CXC chemokine ligand 10. CCL-2: C–C Motif Chemokine ligand 2. IRI: ischemia/reperfusion injury. Two lanes in a row are from the same mixed samples, the in vitro experiment was repeated independently at least 3 times.