seh  (Proteintech)

Journal: Advanced Science

Article Title: CYP4X1/sEH‐Dependent Endocannabinoid Metabolism Drives Fibroblast‐Mediated Immunosuppression to Limit Immunotherapy in Colon Cancer

doi: 10.1002/advs.202507695

Figure Lengend Snippet: CYP4X1 and sEH jointly predict prognosis and immune infiltration in colon cancer. A) Venn diagram showing the endocannabinoid metabolism‐related differentially expressed genes (DEGs) in the TCGA colon adenocarcinoma (TCGA‐COAD) and colon cancer‐related GEO datasets ( GSE39582 and GSE44076 ). B) CYP4X1 and EPHX2 gene expression levels in human colon cancer tissues and normal tissues from the TCGA and GEO databases ( GSE39582 and GSE44076 ). C) Representative immunohistochemistry (IHC) images and IHC scores of CYP4X1 and sEH staining in human colon carcinoma tissue microarrays ( n = 90). Scale bar, 500 µm. D) Kaplan‐Meier survival curves of overall survival for patients with colon cancer in the high CYP4X1 and low sEH expression (CYP4X1 Hi sEH Lo ), high CYP4X1 and high sEH expression (CYP4X1 Hi sEH Hi ), low CYP4X1 and low sEH expression (CYP4X1 Lo sEH Lo ), and low CYP4X1 and high sEH expression (CYP4X1 Lo sEH Hi ) groups stratified by the median expression of CYP4X1 and sEH. E) Representative images of multiplex immunofluorescence (mIF) staining among four groups stratified by the median expression of CYP4X1 and sEH. Scale bar, 50 µm. F) Quantification of CD8 + T cells, FOXP3 + Tregs, and α‐SMA + CAFs as a proportion of total cells ( n = 8). Data are shown as mean ± SEM. P values were determined using Mann‐Whitney tests (B), Wilcoxon matched‐pairs signed‐rank tests (C), log‐rank test (D), or one‐way ANOVA (F). ** P < 0.01; *** P < 0.001.

Article Snippet: Next, slides were incubated with primary antibodies against CYP4X1 (1:100, Invitrogen, #PA5‐101319), sEH (1:200, Proteintech, #10833‐1‐AP), GPR119 (1:50, Affinity, #DF4892), α‐SMA (1:300, BOSTER, #BM0002), FOXP3 (1:400, Cell Signaling Technology, #12653), CD31 (1:300, Santa Cruz, #sc‐376764), CD8 (1:2, DAKO, #IR623), cytokeratin (CK, 1:2, DAKO, #IS05330‐2), PDPN (1:200, Huabio, #ET1703‐61), HLA‐DRA (1:1000, Huabio, #ET1610‐66), or CXCL12 (1:100, Huabio, #ER1902‐35).

Techniques: Gene Expression, Immunohistochemistry, Staining, Expressing, Multiplex Assay, Immunofluorescence, MANN-WHITNEY

Journal: Advanced Science

Article Title: CYP4X1/sEH‐Dependent Endocannabinoid Metabolism Drives Fibroblast‐Mediated Immunosuppression to Limit Immunotherapy in Colon Cancer

doi: 10.1002/advs.202507695

Figure Lengend Snippet: CYP4X1 and EPHX2 manipulate the immune microenvironment in murine colon cancer models. A) Biochemistry of the CYP4X1‐sEH pathway. AEA, anandamide; 14,15‐EET‐EA, 14,15‐epoxyeicosatrienoic acid‐ethanolamide; 14,15‐DiHET‐EA, 14,15‐dihydroxyeicosatrienoic acid‐ethanolamide. B) MC38 tumor chunks with Cyp4x1 knockdown ( Cyp4x1 KD ), Ephx2 overexpression ( Ephx2 OE ), or their combination ( Cyp4x1 KD Ephx2 OE ) were orthotopically implanted into C57BL/6 mice ( n = 5). 14,15‐EET‐EA level was determined by liquid chromatography tandem‐mass spectrometry (LC‐MS/MS). 14,15‐EET‐EA level was reported as pmol mg −1 of wet tissue weight. C) Representative ex vivo images of orthotopic MC38 tumors and tumor weights. D) Representative flow staining and quantification of CD8 + T cells, CD4 + T cells, Tregs, CD107a + CD8 + T cells, and IFN‐γ + CD8 + T cells in tumor tissues of the indicated groups. E) IHC staining and quantification of CD8 and FOXP3 in the indicated tumor tissues. Scale bar, 50 µm. F) Representative images and quantitative analysis of immunofluorescence (IF) staining of CD8 (red) and GZMB (green) or IFN‐γ (green) in tumor tissues. Scale bar, 50 µm. G) CT26 colon cancer cells with Cyp4x1 KD , Ephx2 OE , or Cyp4x1 KD Ephx2 OE expression were subcutaneously implanted into BALB/c mice ( n = 8). Tumor weights and tumor volumes were presented. H) 14,15‐EET‐EA level was determined by LC‐MS/MS. Data are presented as mean ± SEM. P values were determined using one‐way ANOVA. * P < 0.05 and ** P < 0.01 vs. control; ^ P < 0.05 and ^^ P < 0.01 vs. Cyp4x1 KD ; # P < 0.05 and ## P < 0.01 vs. Ephx2 OE group.

Article Snippet: Next, slides were incubated with primary antibodies against CYP4X1 (1:100, Invitrogen, #PA5‐101319), sEH (1:200, Proteintech, #10833‐1‐AP), GPR119 (1:50, Affinity, #DF4892), α‐SMA (1:300, BOSTER, #BM0002), FOXP3 (1:400, Cell Signaling Technology, #12653), CD31 (1:300, Santa Cruz, #sc‐376764), CD8 (1:2, DAKO, #IR623), cytokeratin (CK, 1:2, DAKO, #IS05330‐2), PDPN (1:200, Huabio, #ET1703‐61), HLA‐DRA (1:1000, Huabio, #ET1610‐66), or CXCL12 (1:100, Huabio, #ER1902‐35).

Techniques: Knockdown, Over Expression, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Ex Vivo, Staining, Immunohistochemistry, Immunofluorescence, Expressing, Control

Journal: Advanced Science

Article Title: CYP4X1/sEH‐Dependent Endocannabinoid Metabolism Drives Fibroblast‐Mediated Immunosuppression to Limit Immunotherapy in Colon Cancer

doi: 10.1002/advs.202507695

Figure Lengend Snippet: CYP4X1/sEH‐derived 14,15‐EET‐EA mediates tumor immunosuppression via upregulation of PD‐L1, CXCL12, and TGF‐β in CAFs. A) The production of 14,15‐EET‐EA in HCT116 and HT29 colon cancer cells expressing CYP4X1 KD , EPHX2 OE , or their combination ( CYP4X1 KD EPHX2 OE ) was measured by LC‐MS/MS ( n = 5). MRC5 fibroblasts were incubated with 14,15‐EET‐EA or conditioned medium (CM) from HCT116 or HT29 cells of the indicated groups ( n = 5). B) αSMA and FAP mRNA expression levels in MRC5 fibroblasts incubated with 14,15‐EET‐EA or CM from HCT116 or HT29 cells of the indicated groups were measured by qPCR. C) Transwell migration analysis of Tregs cocultured with the above‐treated MRC5 fibroblasts. D) Transwell migration analysis of endothelial cells (HUVECs) and pericyte cells (human brain vascular pericytes) cocultured with the CM from above‐treated MRC5 fibroblasts. E) Carboxyfluorescein succinimidyl ester (CFSE) dilution was used to assess the proliferation of CD8 + T cells cocultured with the above‐treated MRC5 fibroblasts. F) The percentages of CD107a + CD8 + T cells and IFN‐γ + CD8 + T cells were determined by flow cytometry. G) CXCL12 and TGF‐β production in MRC5 fibroblasts of the indicated groups was measured by ELISA. H) PD‐L1 expression level in CAFs was determined by flow cytometry. MRC5 fibroblasts pretreated with 14,15‐EET‐EA were incubated with the treatment of alone or a combination (Abs) of neutralizing anti‐PD‐L1, anti‐CXCL12, and anti‐TGF‐β antibodies ( n = 3). I) Transwell migration analysis of Tregs cocultured with the above‐treated MRC5 fibroblasts. J) CFSE dilution was used to measure the proliferation of CD8 + T cells cocultured with the above‐treated MRC5 fibroblasts. K) GZMB and IFN‐γ mRNA levels in CD8 + T cells were determined by qPCR. L) Transwell migration analysis of endothelial cells cocultured with the CM from the above‐treated MRC5 fibroblasts. Data are presented as mean ± SEM. P values were determined using one‐way ANOVA. & P < 0.05 and && P < 0.01 vs. control; * P < 0.05 and ** P < 0.01 vs. vector; ^ P < 0.05 and ^^ P < 0.01 vs. CYP4X1 KD ; # P < 0.05 and ## P < 0.01 vs. EPHX2 OE ; Δ P < 0.05 and ΔΔ P < 0.01 vs. 14,15‐EET‐EA group.

Article Snippet: Next, slides were incubated with primary antibodies against CYP4X1 (1:100, Invitrogen, #PA5‐101319), sEH (1:200, Proteintech, #10833‐1‐AP), GPR119 (1:50, Affinity, #DF4892), α‐SMA (1:300, BOSTER, #BM0002), FOXP3 (1:400, Cell Signaling Technology, #12653), CD31 (1:300, Santa Cruz, #sc‐376764), CD8 (1:2, DAKO, #IR623), cytokeratin (CK, 1:2, DAKO, #IS05330‐2), PDPN (1:200, Huabio, #ET1703‐61), HLA‐DRA (1:1000, Huabio, #ET1610‐66), or CXCL12 (1:100, Huabio, #ER1902‐35).

Techniques: Derivative Assay, Expressing, Liquid Chromatography with Mass Spectroscopy, Incubation, Migration, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Control, Plasmid Preparation

Journal: Advanced Science

Article Title: CYP4X1/sEH‐Dependent Endocannabinoid Metabolism Drives Fibroblast‐Mediated Immunosuppression to Limit Immunotherapy in Colon Cancer

doi: 10.1002/advs.202507695

Figure Lengend Snippet: CYP4X1/sEH‐derived 14,15‐EET‐EA triggers immunosuppression via GPR119‐Gs/β‐arrestin 2 signaling. MRC5 fibroblasts were treated with or without GPR119 knockdown, followed by stimulation with 14,15‐EET‐EA. A) αSMA, PD‐L1, CXCL12, and TGF‐β mRNA levels in the above‐treated MRC5 fibroblasts ( n = 5). B) Transwell migration analysis of Tregs cocultured with the above‐treated MRC5 fibroblasts ( n = 5). C) GZMB and IFNγ mRNA levels in CD8 + T cells cocultured with the above‐treated MRC5 fibroblasts were measured by qPCR ( n = 5). D) MC38‐luciferase cells were mixed with Gpr119 shRNA‐ or scramble shRNA‐treated L929 fibroblasts, and then co‐implanted into C57BL/6 mice. In vivo bioluminescent images and quantification of the indicated groups ( n = 8). E) Representative IF staining of αSMA (red) and vimentin (green) in MC38 tumor tissues ( n = 5). Scale bar, 50 µm. F) PD‐L1, CXCL12, and TGF‐β mRNA levels in the CAFs ( n = 5). G) The infiltration of CD8 + T cells in tumor tissues was determined by flow cytometry ( n = 5). H) Representative IF staining and frequency analysis of CD8 (red) and GZMB (green) or IFN‐γ (green) in tumor tissues ( n = 5). Scale bar, 50 µm. I) Treg accumulation in tumor tissues was analyzed and quantified by flow cytometry ( n = 5). J) IF analysis of tumor vessel normalization in the indicated groups ( n = 5). Scale bar, 50 µm. K) cAMP level in MRC5 and L929 fibroblasts incubated with the CM from CYP4X1 KD EPHX2 OE ‐expressed colon cancer cells with or without 14,15‐EET‐EA supplementation ( n = 5). L,M) β‐arrestin 2, p‐EGFR, and p‐Akt protein levels in MRC5 and L929 fibroblasts incubated with CM from CYP4X1 KD EPHX2 OE ‐expressed colon cancer cells with or without 14,15‐EET‐EA supplementation ( n = 5). N) β‐arrestin 2, p‐EGFR, and p‐Akt protein levels in L929 and MRC5 fibroblasts with or without GPR119 knockdown, followed by stimulation with 14,15‐EET‐EA. O,P) p‐STAT3 protein level, PD‐L1, CXCL12, and TGF‐β mRNA levels in MRC5 fibroblasts of the indicated groups ( n = 5). Data are presented as mean ± SEM. P values were determined using one‐way ANOVA (A‐C, K‐M, and P) or Student's t ‐tests (D, F‐J). * P < 0.05 and ** P < 0.01 vs. control or sh‐Scram; ^ P < 0.05 and ^^ P < 0.01 vs. 14,15‐EET‐EA+si‐Scram or 14,15‐EET‐EA; # P < 0.05 and ## P < 0.01 vs. CYP4X1 KD EPHX2 OE group.

Article Snippet: Next, slides were incubated with primary antibodies against CYP4X1 (1:100, Invitrogen, #PA5‐101319), sEH (1:200, Proteintech, #10833‐1‐AP), GPR119 (1:50, Affinity, #DF4892), α‐SMA (1:300, BOSTER, #BM0002), FOXP3 (1:400, Cell Signaling Technology, #12653), CD31 (1:300, Santa Cruz, #sc‐376764), CD8 (1:2, DAKO, #IR623), cytokeratin (CK, 1:2, DAKO, #IS05330‐2), PDPN (1:200, Huabio, #ET1703‐61), HLA‐DRA (1:1000, Huabio, #ET1610‐66), or CXCL12 (1:100, Huabio, #ER1902‐35).

Techniques: Derivative Assay, Knockdown, Migration, Luciferase, shRNA, In Vivo, Staining, Flow Cytometry, Incubation, Control

Journal: Advanced Science

Article Title: CYP4X1/sEH‐Dependent Endocannabinoid Metabolism Drives Fibroblast‐Mediated Immunosuppression to Limit Immunotherapy in Colon Cancer

doi: 10.1002/advs.202507695

Figure Lengend Snippet: The CYP4X1/sEH‐14,15‐EET‐EA‐GPR119 axis regulates sensitivity to anti‐PD‐1 therapy. A) MC38 colon cancer cells with or without Cyp4x1 KD Ephx2 OE expression were subcutaneously implanted into C57BL/6 mice and treated with IgG or anti‐PD‐1 antibody (α‐PD‐1). Tumor growth curves (Two‐way ANOVA) and tumor weights were presented ( n = 8). B) 14,15‐EET‐EA level was determined by LC‐MS/MS ( n = 6). C) Representative flow staining and quantification of CD107a + CD8 + T cells and IFN‐γ + CD8 + T cells in tumor tissues of the indicated groups ( n = 6). D) Representative IF staining and quantification of CD8 (red) and GZMB (green) or IFN‐γ (green) in tumor tissues ( n = 6). Scale bar, 50 µm. E) Percentages of CD8 + T cells and Tregs ( n = 6). F) Tumor growth curves (Two‐way ANOVA) and tumor burdens of MC38 tumor‐bearing mice treated with IgG control, anti‐PD‐1 alone, or in combination with 14,15‐EET‐EA ( n = 8). G) C57BL/6 mice were co‐implanted with MC38 cells and Gpr119 shRNA‐ or scramble shRNA‐treated L929 fibroblasts, followed by anti‐PD‐1 treatment. Tumor weights were measured in the indicated groups ( n = 9). H) C57BL/6 mice inoculated with MC38 cells were treated with GPR119 agonist AR231453 , anti‐PD‐1 alone, or their combination. Tumor weights were measured in the indicated groups ( n = 7). I) Representative flow staining and quantification of CD8 + T cells, CD107a + CD8 + T cells, and IFN‐γ + CD8 + T cells in tumor tissues of the indicated groups ( n = 5). J) Treg accumulation in tumors was analyzed and quantified by IHC staining ( n = 5). Data are presented as mean ± SEM. P values were determined using one‐way ANOVA (A‐D, F‐I) and Student's t ‐tests (E and J). * P < 0.05 and ** P < 0.01 vs. control; # P < 0.05 and ## P < 0.01 vs. α‐PD‐1 group.

Article Snippet: Next, slides were incubated with primary antibodies against CYP4X1 (1:100, Invitrogen, #PA5‐101319), sEH (1:200, Proteintech, #10833‐1‐AP), GPR119 (1:50, Affinity, #DF4892), α‐SMA (1:300, BOSTER, #BM0002), FOXP3 (1:400, Cell Signaling Technology, #12653), CD31 (1:300, Santa Cruz, #sc‐376764), CD8 (1:2, DAKO, #IR623), cytokeratin (CK, 1:2, DAKO, #IS05330‐2), PDPN (1:200, Huabio, #ET1703‐61), HLA‐DRA (1:1000, Huabio, #ET1610‐66), or CXCL12 (1:100, Huabio, #ER1902‐35).

Techniques: Expressing, Liquid Chromatography with Mass Spectroscopy, Staining, Control, shRNA, Immunohistochemistry

Journal: Advanced Science

Article Title: CYP4X1/sEH‐Dependent Endocannabinoid Metabolism Drives Fibroblast‐Mediated Immunosuppression to Limit Immunotherapy in Colon Cancer

doi: 10.1002/advs.202507695

Figure Lengend Snippet: CYP4X1/sEH‐GPR119 axis predicts the response to anti‐PD‐1 therapy in colon cancer. A) Immunophenoscore in the CYP4X1 Hi vs. CYP4X1 Lo groups, EPHX2 Hi vs. EPHX2 Lo groups, and the GPR119 Hi and GPR119 Lo groups ( GSE33193 ). B‐D) Kaplan‐Meier plots of overall survival and progression‐free survival for patients receiving PD‐1 blockade therapy according to CYP4X1 , EPHX2 , or GPR119 expression. E) Representative images of mIF staining in human colon cancer tissues with low GPR119 + CAF or high GPR119 + CAF abundance. Scale bar, 50 µm. F) Quantification of CD8 + T cells and FOXP3 + Tregs as a proportion of total cells ( n = 6). G,H) Representative images of IHC staining and quantification of CYP4X1 and sEH in microsatellite stability (MSS; n = 15) and microsatellite instability‐high (MSI‐H; n = 6) human colon cancer tissues. Scale bar, 50 µm. I) Representative IF staining and quantification of αSMA (red) and GPR119 (green) in MSS ( n = 15) and MSI‐H ( n = 6) human colon cancer tissues. Scale bar, 50 µm. Data are shown as mean ± SEM. P values were determined using Mann‐Whitney tests (A), log‐rank tests (B‐D), or Student's t ‐tests (F, H, and I). ** P < 0.01.

Article Snippet: Next, slides were incubated with primary antibodies against CYP4X1 (1:100, Invitrogen, #PA5‐101319), sEH (1:200, Proteintech, #10833‐1‐AP), GPR119 (1:50, Affinity, #DF4892), α‐SMA (1:300, BOSTER, #BM0002), FOXP3 (1:400, Cell Signaling Technology, #12653), CD31 (1:300, Santa Cruz, #sc‐376764), CD8 (1:2, DAKO, #IR623), cytokeratin (CK, 1:2, DAKO, #IS05330‐2), PDPN (1:200, Huabio, #ET1703‐61), HLA‐DRA (1:1000, Huabio, #ET1610‐66), or CXCL12 (1:100, Huabio, #ER1902‐35).

Techniques: Expressing, Staining, Immunohistochemistry, MANN-WHITNEY

Journal: Redox Biology

Article Title: Endothelial soluble epoxide hydrolase links polyunsaturated fatty acid metabolism to oxidative stress and atherosclerosis progression

doi: 10.1016/j.redox.2025.103730

Figure Lengend Snippet: Atheroprone endothelial cells enriched with genes of polyunsaturated fatty acid metabolism. A. tSNE plots showing endothelial cell clustering from murine aortae following scRNA sequencing originally published in the Kalluri et al., 2019 and Kan et al., 2021. B. Bubble plots showing key endothelial cell (EC) markers of the clusters presented in panel A. C. Atherosclerosis and oscillatory shear stress (OSS) gene score enrichment in EC1 and EC2 presented in panel A. D. Glycolysis, tricarboxylic acid (TCA) and fatty acid oxidation (FAO) gene score enrichment in EC1 and EC2 presented in panel A. E. Polyunsaturated fatty acid (PUFA) and Cyp450 metabolism gene score enrichment in EC1 and EC2 presented in panel A. F. Heatmaps showing the top 25 marker genes enriched in EC clusters presented in panel A. G. tSNE plots showing Ephx2 expression in the clusters presented in panel A. Red dotted cycles indicating EC1 cluster as in panel A.

Article Snippet: Generation of floxed sEH ( Ephx2 fl/fl ) mice was performed by TaconicArtemis GmbH (Cologne, Germany) as previously described [ ].

Techniques: Sequencing, Shear, Marker, Expressing

Journal: Redox Biology

Article Title: Endothelial soluble epoxide hydrolase links polyunsaturated fatty acid metabolism to oxidative stress and atherosclerosis progression

doi: 10.1016/j.redox.2025.103730

Figure Lengend Snippet: Endothelial soluble epoxide hydrolase accelerates atherosclerosis development. A. Representative brightfield image and oil-red-o staining of left carotid artery from wild type and inducible endothelial-specific sEH knockout mice (iΔEC) following 4 weeks of AAV-PCSK9, 4 weeks of HFD and 3 weeks of partial carotid ligation. Quantification of plaque area is shown on the right. Bar = 1 mm and 200 μm in brightfield image and sections, respectively. n = 6 mice/group, two-way ANOVA. B. Representative brightfield image and oil-red-o staining of left carotid artery from wild type and inducible endothelial-specific sEH overexpression mice (iOE) following 4 weeks of AAV-PCSK9, 4 weeks of HFD and 3 weeks of partial carotid ligation. Quantification of plaque area is shown on the right. Bar = 1 mm and 200 μm in brightfield image and sections, respectively. n = 6 mice/group, two-way ANOVA. C. Representative immunefluorescent image of VCAM-1 (green), CD31 (red) and nuclei (DAPI) from left carotid sections of mice as in panel B. Bar = 50 μm. n = 6 mice/group, Student's t-test. D. Representative immunofluorescent image of F4/80 (green), CD31 (red) and nuclei (DAPI) from left carotid sections of mice as in panel B. Bar = 50 μm. n = 6 mice/group, Student's t-test.

Article Snippet: sEH-tdTomato reporter mice ( Ephx2 LSL-tdTomato ) and sEH overexpression ( Rosa26 CAG-LSL-Ephx2−3xFlag ) were generated by Shanghai Model Organisms Center (Shanghai, China) using CRISPR/Cas9 system in a C57BL/6J mouse background.

Techniques: Staining, Knock-Out, Ligation, Over Expression

Journal: Redox Biology

Article Title: Endothelial soluble epoxide hydrolase links polyunsaturated fatty acid metabolism to oxidative stress and atherosclerosis progression

doi: 10.1016/j.redox.2025.103730

Figure Lengend Snippet: Re-establishing redox balance suppresses sEH mediated TGFβ activation and atherosclerosis development. A. Volcano plot showing differentially regulated transcripts in endothelial cells isolated from wild type or sEH overexpressing mice. n = 3 independent cell batches/group. B. Transcriptional factor activity prediction in samples as in panel A. C. Representative immunoblot showing protein levels in human endothelial cells infected with a control (CTL) or a lentivirus overexpressing sEH and treated with solvent or N-acetyl cysteine (NAC, 1 mmol/L, 24 h). n = 5 independent cell batches/group, two-way ANOVA. D. Relative mRNA levles of TGFβ and its targets in samples as in panel C. n = 4 independent cell batches/group. E. Representative MitoSOX fluorescence in cells as in panel C. similar observations were made in 5 more independent cell batches/group F. Relative levels of MitoSOX fluorescent intensity following flow cytometry in cells as in panel C. n = 6 independent cell batches/group, two-way ANOVA. G. Representative brightfield image and oil-red-o staining of left carotid artery from endothelial cell specific sEH inducible overexpression mice (OE) treated with solvent (Sol) or N-acetyl cysteine (NAC), following 4 weeks of AAV-PCSK9, 4 weeks of HFD and 3 weeks of partial carotid ligation. Quantification of plaque area is shown on the right. Bar = 1 mm and 200 μm in brightfield image and sections, respectively. n = 4 mice/group, two-way ANOVA. H. Representative immunofluorescent image of pSMAD2 (red), CD31 (cyan) and DAPI (green) on left carotid artery in mice as in panel G. Quantification of pSMAD2 levels in endothelial cells is shown on the right. Bar = 20 μm. n = 4–5 mice/group, one-way ANOVA.

Article Snippet: sEH-tdTomato reporter mice ( Ephx2 LSL-tdTomato ) and sEH overexpression ( Rosa26 CAG-LSL-Ephx2−3xFlag ) were generated by Shanghai Model Organisms Center (Shanghai, China) using CRISPR/Cas9 system in a C57BL/6J mouse background.

Techniques: Activation Assay, Isolation, Activity Assay, Western Blot, Infection, Control, Solvent, Fluorescence, Flow Cytometry, Staining, Over Expression, Ligation