Fig. 5

AKBA may activate U-STAT1 expression and promote BST2 transcription by anchoring BST2. (A) Expression of BST2 in peripheral blood of patients with septic shock. (B) Expression of ISG15, BST2, OAS2, IFIT1, IFIT3 and STAT1 in LPS + AKBA group, n = 3. (C) Molecular docking results and site prediction of AKBA and BST2. (D) Top 20 items for differential protein enrichment analysis and a special concentration of STAT1 signaling pathway. (E, F) The interaction between AKBA and BST2 was evaluated by CETSA technique and CETSA dissolution curve, n = 3. (G) mRNA expression levels of STAT1 and BST2 in HPMECs were evaluated using reverse transcription-quantitative PCR after regulation of STAT1 or BST2, n = 3. (H) Relative luciferase activity was assessed using the dual-luciferase reporter assay in HPMECs co-transfected with OE-STAT1 or OE-NC and BST2-WT or BST2-MUT. (I, J) The molecular dynamics simulation of AKBA and BST2 complex system was conducted to investigate the binding stability and kinetic behavior. (K, L) DARTS-WB assesses the stability of AKBA and BST2 complex systems. (M, N) CHlP-gPCR was performed in HPMECs transfected with pcDNA3.1-HA-AP to identify the enrichment of HA-STAT1 onto BST2 promoter region, IgG served as an antibody control. n = 3. ^*p < 0.05, ^**p < 0.01 and ^***p < 0.001.