Fig 2: ATF6 regulates TRIM10 transcription in HUVECs. (A) The expression of TRIM10 after stimulation with 10 μM Ceapin‐A7 or AA147 and 10 μg/mL LPS was determined by RT‐qPCR (*p < 0.05, N = 3). (B) The expression of TRIM10 after ATF6 knockout or ATF6 overexpression and stimulation with 10 μg/mL LPS was determined by RT‐qPCR (*p < 0.05, N = 3). (C) Expression and semiquantitative analysis of TRIM10 after 10 μM Ceapin‐A7 or AA147 treatment and stimulation with 10 μg/mL LPS by western blot (*p < 0.05, N = 3); (D) western blot analysis of the expression and semiquantitative analysis of TRIM10 after ATF6 knockout/overexpression in cells stimulated with 10 μg/mL LPS (*p < 0.05, N = 3). (E) Western blot analysis of the expression and semiquantitative analysis of TRIM10 and phospho‐NF‐κB p65 in ATF6‐overexpressing HUVECs after 10 μM Ceapin‐A7 stimulation with 10 μg/mL LPS (*p < 0.05, N = 3); (F) gene expression of TNF‐α and IL‐6 in ATF6‐overexpressing HUVECs after 10 μM Ceapin‐A7 stimulation with 10 μg/mL LPS, as determined by RT–qPCR (*p < 0.05, N = 3). (G) The putative ATF6α:NF‐YA binding sites in the TRIM10 promoter and the nucleotide sequences representing the predicted binding sequences are shown, with the red letters indicating core elements. (H) Luciferase activity of transfected HUVECs transfected with TRIM10 and its mutant construct after treatment with 10 μg/mL LPS determined by dual‐luciferase reporter assay (*p < 0.05, N = 4). (I) The binding of ATF6α to the TRIM10 promoter in HUVECs after treatment with 10 μg/mL LPS, as determined by ChIP‐qPCR (*p < 0.05, N = 3).

Fig 3: TRIM10 expression following extended hepatectomy (PH80) and major hepatectomy (PH70) and its role in endothelial inflammation. (A) Hepatic TRIM10 expression following PH80 (N = 5) and PH70 (N = 5) in WT mice, PH80 (N = 5) in ATF6‐knockout mice, as well as PH80 (N = 5) in LSEC ATF6‐specific deficiency mice, as determined by immunohistochemistry (original magnification ×400, scale bars: 50 μm) and semiquantitative analysis (*p < 0.05); (B) analysis of hepatic TRIM10 expression following marginal hepatectomy (N = 8) and minor hepatectomy (N = 6) in humans by immunohistochemistry (original magnification ×400, scale bars: 50 μm) and semiquantitative analysis (*p < 0.05). (C) The expression and quantification of TRIM10 in HUVECs with and without TRIM10 siRNA knockdown after stimulation with 10 μg/mL LPS were determined by RT–qPCR (*p < 0.05, N = 4). (D) Western blot analysis of the expression and semiquantitative analysis of TRIM10 and phospho‐NF‐κB p65 in HUVECs with and without TRIM10 siRNA knockdown after stimulation with 10 μg/mL LPS (*p < 0.05, N = 3). (E, F) Gene expression of TNF‐α and IL‐6 in cells with and without TRIM10 siRNA knockdown after stimulation with 10 μg/mL LPS, as determined by RT–qPCR (*p < 0.05, N = 3).

Fig 4: ATF6 suppresses inflammation through the NF‐κB signaling pathway in HUVECs. (A) Gene Set Enrichment Analysis of ATF6 inhibition (10 μM Ceapin‐A7) under 10 mg/mL tunicamycin‐induced UPR in HUVECs revealed that the NF‐κB signaling pathway was activated during ATF6‐mediated inflammation. (B, C) Gene expression of TNF‐α and IL‐6 in WT and ATF6‐knockout HUVECs treated with or without 10 μM Ceapin‐A7 and stimulated with 10 μg/mL LPS determined by RT–qPCR (*p < 0.05, N = 3). (D) Western blot analysis of the expression and semiquantitative analysis of phospho‐NF‐κB p65 and NF‐κB p65 in WT and ATF6‐knockout HUVECs treated with or without 10 μM Ceapin‐A7 after stimulation with 10 μg/mL LPS (*p < 0.05, N = 4). (E) The expression of tripartite motif‐containing protein 10 (TRIM10) and TANK‐binding kinase 1 (TBK1) after ATF6 inhibition with 10 μM Ceapin‐A7 and stimulation with 10 mg/mL tunicamycin was determined by RNA‐seq analysis (*p < 0.05, N = 3). (F) Western blot analysis of the expression and semiquantitative analysis of phospho‐NF‐κB p65 and NF‐κB p65 in WT and ATF6‐overexpressing HUVECs treated with or without 10 μM Ceapin‐A7/AA147 after stimulation with 10 μg/mL LPS (*p < 0.05, N = 3). (G, H) Gene expression of TNF‐α and IL‐6 in WT and ATF6‐overexpressing HUVECs treated with or without 10 μM AA147 and stimulated with 10 μg/mL LPS determined by ELISA (*p < 0.05, N = 3).

Fig 5: ATF6 regulates TRIM10 transcription in HUVECs. (A) The expression of TRIM10 after stimulation with 10 μM Ceapin‐A7 or AA147 and 10 μg/mL LPS was determined by RT‐qPCR (*p < 0.05, N = 3). (B) The expression of TRIM10 after ATF6 knockout or ATF6 overexpression and stimulation with 10 μg/mL LPS was determined by RT‐qPCR (*p < 0.05, N = 3). (C) Expression and semiquantitative analysis of TRIM10 after 10 μM Ceapin‐A7 or AA147 treatment and stimulation with 10 μg/mL LPS by western blot (*p < 0.05, N = 3); (D) western blot analysis of the expression and semiquantitative analysis of TRIM10 after ATF6 knockout/overexpression in cells stimulated with 10 μg/mL LPS (*p < 0.05, N = 3). (E) Western blot analysis of the expression and semiquantitative analysis of TRIM10 and phospho‐NF‐κB p65 in ATF6‐overexpressing HUVECs after 10 μM Ceapin‐A7 stimulation with 10 μg/mL LPS (*p < 0.05, N = 3); (F) gene expression of TNF‐α and IL‐6 in ATF6‐overexpressing HUVECs after 10 μM Ceapin‐A7 stimulation with 10 μg/mL LPS, as determined by RT–qPCR (*p < 0.05, N = 3). (G) The putative ATF6α:NF‐YA binding sites in the TRIM10 promoter and the nucleotide sequences representing the predicted binding sequences are shown, with the red letters indicating core elements. (H) Luciferase activity of transfected HUVECs transfected with TRIM10 and its mutant construct after treatment with 10 μg/mL LPS determined by dual‐luciferase reporter assay (*p < 0.05, N = 4). (I) The binding of ATF6α to the TRIM10 promoter in HUVECs after treatment with 10 μg/mL LPS, as determined by ChIP‐qPCR (*p < 0.05, N = 3).