Journal: bioRxiv

Article Title: “Tumor-to-endothelium mitochondrial transfer licenses endothelial cells for CD8 + T cell recognition via mitochondrial neoantigen presentation”

doi: 10.64898/2026.01.30.702567

Figure Lengend Snippet: (A) Representative immunofluorescence images showing PD-L1 expression in naïve kidney and RENCA tumors (Hoechst, nuclei; PD-L1, red). IF and qPCR quantification of PD-L1 signal is shown on the right. (B) From top to bottom, schematic of RENCA tumor implantation and treatment schedule, longitudinal tumor growth monitoring and endpoint analyses. (C) IFN γ ELISPOT responses of CD8 + T cells to BMDCs pulsed with TAMAs peptides and/or RENCA mitochondrial lysate. Peptide- and lysate-driven responses are shown as the summed response (spot-forming cells, SFC, per indicated number of CD8 + T cells). (D) Flow cytometric quantification of intratumoral CD8 + T cells across treatment groups. Right, correlation between tumor volume and CD8 + T cell abundance. (E) Immunoblot showing perforin expression in tumors across treatment groups (VCL loading control) with densitometric quantification. (F) Flow cytometric analysis of activated CD8 + T cells (CD107a + ) across treatment groups and correlation with tumor volume. Data are shown as mean ± s.e.m. unless otherwise indicated. Statistical comparisons were performed using one-way ANOVA with multiple-comparison correction.

Article Snippet: Housekeeping gene Rn18s (Mm03928990_g1), PDL-1 (Mm03048248_m1), Perforin (Mm00812512_m1), Ang1 (Mm01233546_g1), Ang2 (Mm00657574_s1), VECAD/Cdh5 (Mm00486938_m1), Vcam1 (Mm01320970_m1), Cxcl10 (Mm00445235_m1), Cxcl11 (Mm00444662_m1), Cxcl9 (Mm00434946_m1), Vegfa (Mm00437306_m1), Pdgfb (Mm00440677_m1), Irf3 (Mm00516784_m1), CD8a (Mm01182107_g1), Ifnγ (Mm01168134_m1), Il12a (Mm00434169_m1), Il12b (Mm01288989_m1), and Gzmb (Mm00442837_m1), TNF (Mm00443258_m1), PTEN (Mm00477208_m1), Itgax (Mm00498701_m1), Tnfsf10 (Mm01283606_m1), Tnfrsf1a (Mm00441883_g1).

Techniques: Immunofluorescence, Expressing, Tumor Implantation, Enzyme-linked Immunospot, Western Blot, Control, Comparison

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: IHC analysis of tumor of K19-Wnt1/C2mE mice. ( A ) Ki-67-positive cell rates and the numbers of Iba1-positive macrophages in the tumors were assessed via IHD (control group [n = 5] vs HFD group [n = 4] vs LPS group [n = 3] vs HFD + LPS group [n = 6]; bar = 100 μm). A magnified view (× 400) of the area is shown on the left side. The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was performed via the Tukey‒Kramer test. ( B ) Representative images of CD3-, CD4-, CD8-, and FOXP3-positive lymphocytes in the tumor stroma with the numbers of them per HPFs (control group [n = 4] vs HFD group [n = 4] vs LPS group [n = 3] vs HFD + LPS group [n = 5]; bar = 100 μm). A magnified view (×400) of the area is shown on the left side. The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was performed via the Tukey‒Kramer test. ( C ) Representative images of cytoplasmic expressions of TNF-α, PD-L1, and PD-1 in the tumor with their IHC scores (control group [n = 5] vs HFD group [n = 4] vs LPS group [n = 3] vs HFD + LPS group [n = 5]; bar = 100 μm). A magnified view (×400) of the area is shown on the left side. The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis for the TNF-α and PD-L1 score and the PD-1-positive cell numbers were performed via the Steel-Dwass test and the Tukey–Kramer test, respectively. ( D ) Representative image of dual-staining of TNF-α ( blue ) and F4/80 ( brown ) for tumors from the HFD + LPS group of mice. An enlarged view of the area with double staining is shown on the lower side. Cells with dual staining are indicated by black arrows . Bar = 50 μm. Ctrl, control.

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: Control, Staining, Double Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: Alteration in signaling pathways related to tumor growth and immune evasion in the HFD + LPS group of K19-Wnt1/C2mE mice. ( A ) The 8-OHdG content ( left ) and Nqo1 mRNA/ Gapdh mRNA ratio ( right ) of the tumors (control group [n = 4] vs HFD group [n = 4] vs LPS group [n = 3] vs HFD + LPS group [n = 6]). The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. ( B ) Representative images of western blot analysis of TNF-α, TLR4, PD-L1, p-NFκB p65, NFκB, and GAPDH in tumors. The results represent at least 3 independent experiments. ( C ) The density ratio of each band to GAPDH of Western blot analysis was calculated using Image Lab software (Bio-Rad), and the results are presented as box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. ( D ) The relative gene expression levels of Cxcl9 , Ifng , Tnfa , and Cd274 to Gapdh ( Cxcl9, Ifng, Tnf, Cd274 : control group [n = 4] vs HFD group [n = 4] vs LPS group [n = 3] vs HFD + LPS group [n = 6]). The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. Ctrl, control.

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: Protein-Protein interactions, Control, Western Blot, Software, Gene Expression

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: Effects of macrophage depletion on signaling pathways in K19-Wnt1/C2mE mice. ( A ) The relative gene expression levels of Cxcl9 , Ifng , Tnfa, and Cd274 to Gapdh in tumor in the control group, HFD + LPS group, and HFD + LPS + CL group ( Cxcl9, Ifng, Tnfa, Cd274 : control group [n = 5] vs HFD + LPS group [n = 5] vs HFD + LPS + CL group [n = 4]). The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was performed via the Tukey‒Kramer test. ( B ) Representative images of Western blot analysis of the TLR4, PD-L1, p-NFκB p65, and NFκB proteins in the tumors in the control group, HFD + LPS group, and HFD + LPS + CL group. The results represent 3 independent experiments. ( C ) The density ratio of each band to GAPDH of Western blot analysis was calculated using Image Lab software (Bio-Rad), and the results are presented as box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. Ctrl, control.

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: Protein-Protein interactions, Gene Expression, Control, Western Blot, Software

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: In vitro study using MKN7 cells and THP-1-derived macrophages. ( A, B ) Western blot analysis of the TLR4, TNFR1, PD-L1, p-NFκB p65, and NFκB proteins in MKN7 cells stimulated with or without E coli LPS or TNF-α. The results represent 4 independent experiments. The value of the density ratio of each band to that of β-actin was calculated via Image Lab (Bio-Rad), and the data are shown in box plots in B (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. ( C, D ) Effects of BAY 11-7082, an NFκB inhibitor, on PD-L1, p-NFκB p65, and NFκB protein expression in MKN7 cells, as shown by Western blot analysis. The results represent 3 independent experiments. The value of the density ratio of each band to that of β-actin was calculated via Image Lab, and the data are shown in box plots in D (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. ( E ) Representative microscopy image of THP-1-derived macrophages (bar = 200 μm). ( F, G ) Representative images of Western blot analysis of F4/80, CD163, TLR4, TNF-α, and IFN-γ protein expression in PMA-treated THP-1 cells stimulated with E coli LPS. The results represent 3 independent experiments. The value of the density ratio of each band to that of β-actin was calculated via Image Lab, and the data are shown in box plots in G (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via Student’s t- test. BAY, BAY 11-7082; Ctrl, control.

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: In Vitro, Derivative Assay, Western Blot, Expressing, Microscopy, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: In vitro study using the coculture system of MKN7 cells and THP-1-derived macrophages. ( A ) Schematic of the coculture system of MKN7 cells with THP-1-derived macrophages with or without pretreatment with E coli LPS. ( B ) TNF-α concentration in the supernatant of the coculture system. (Control [n = 6] vs E coli LPS + human IgG1 antibody [n = 6] vs E coli LPS + TNF-α-neutralizing antibody [n = 6]). The data are represented in box plots (x = mean). ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. ( C ) Protein expression of PD-L1, p-NFκB p65, and NFκB in MKN7 cells cocultured with E coli LPS-prestimulated macrophages treated with a TNF-α-neutralizing antibody or a human IgG1 antibody in the medium. The results represent 3 independent experiments. ( D ) The density ratio of each band to β-actin of Western blot analysis was calculated using Image Lab software (Bio-Rad), and the results are presented as box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. Ctrl, control.

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: In Vitro, Derivative Assay, Concentration Assay, Control, Expressing, Western Blot, Software

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: Decrease in expressions of inflammatory cytokines and PD-L1 in Nrf2 -KO K19-Wnt1/C2mE mice stimulated with HFD + LPS. ( A ) The relative gene expression levels of and the relative gene expression levels of Nqo1 Cxcl9 , Ifng , Tnfa, and Cd274 to Gapdh in the tumors (K19-Wnt1/C2mE mice control group [n = 4] vs K19-Wnt1/C2mE mice HFD + LPS group [n = 4] vs Nrf2 -KO-K19-Wnt1/C2mE mice control group [n = 4] vs Nrf2 -KO-K19-Wnt1/C2mE mice HFD + LPS group [n = 6]). The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. ( B ) Representative photos of western blot analysis of protein expression of TLR4, PD-L1, p-NFκB p65, and NFκB in the tumors. The results represent 4 independent experiments. ( C ) The density ratio of each band to GAPDH of Western blot analysis was calculated using Image Lab software (Bio-Rad), and the results are presented as box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. Ctrl, control.

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: Gene Expression, Control, Western Blot, Expressing, Software

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: A role of NRF2 stress response in PD-L1 induction by LPS stimulation in MKN7 cells. ( A ) ROS activity in LPS- or TNF-α-stimulated MKN7 cells (control [n = 7] vs LPS [n = 7] vs TNF-α [n = 7]. The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was performed via the Tukey‒Kramer test. ( B ) Representative images of Western blot analysis of cytoplasmic ( left upper ) and nuclear ( left lower ) NRF2 protein expression in MKN7 cells stimulated with LPS. The results represent 3 independent experiments. The value of the density ratio of each band to that of b-actin ( right upper ) or histone H3 ( right lower ) was calculated via Image Lab (Bio-Rad), and the data are shown in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was conducted via Student’s t- test. ( C ) The relative gene expression levels of Nqo1 ( left ) and Cd274 ( right ) to Gapdh in MKN7 cells stimulated with LPS (control [n = 3] vs LPS [n = 3]). The data are represented in box plots (x = mean). ∗ P < .05, ∗∗ P < .01. Statistical analysis was performed via Student’s t- test. ( D ) The relative gene expression levels of NEF2L2 ( left ), NQO1 ( middle ), and CD274 ( right ) to GAPDH in Nrf2 siRNA-transfected MKN7 cells or control siRNA-transfected cells under LPS stimulation (control siRNA + LPS group [n = 3] vs NRF2 siRNA + LPS group [n = 3]). The data are represented in box plots (x = mean). ∗∗ P < .01. Statistical analysis was performed via Student’s t- test. ( E ) ChIP assay using an anti-NRF2 antibody in MKN7 cells treated with or without E coli LPS. Comparison of NRF2 binding to the CD274 promoter (control [n = 3] vs LPS [n = 3]). The values are represented in box plots (x = mean). ∗∗ P < .01. Statistical analysis was performed via Student’s t- test. Ctrl, control.

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: Activity Assay, Control, Western Blot, Expressing, Gene Expression, Transfection, Comparison, Binding Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: Association among protein expression of NQO1 and PD-L1, and Iba1-positive macrophage infiltration in tumor stroma in IHC study in patients with GCA with or without MetS. ( A ) Representative immunohistochemical images of Iba1 ( upper ), NQO1 ( middle ), and PD-L1 ( lower ) in patients with GCA with or without MetS. The cytoplasmic expression of Iba1 in stromal inflammatory cells, and that of NQO1 and PD-L1 in tumor cells in the patients with GCA without MetS ( left ) and with MetS ( right ). Bar = 100 μm. ( B ) The numbers of Iba1-positive macrophage infiltration ( left ), the NQO1 index ( middle ), and the PD-L1 index ( right ) in patients with MetS were significantly higher than those without. The data are represented in box plots (n = 10). ∗ P < .05, ∗∗ P < .01. Student’s t -test and Wilcoxon rank-sum test were applied for statistical analysis of the Iba1-positive cells numbers and the NQO1 and PD-L1 index, respectively. ( C ) Spearman‘s correlation analysis demonstrated positive correlations between the PD-L1 index and the degree of macrophage infiltration and between the PD-L1 index and the NQO1 index (Spearman`s rank correlation coefficient (ρ) value: Iba1 vs PD-L1: ρ = 0.54, P = .013; NQO1 vs PD-L1: ρ = 0.76, P = .0001).

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: Expressing, Immunohistochemical staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Metabolic Syndrome Develops Cardia Cancer via Nuclear Factor-E2-related Factor 2-Programmed Death-Ligand 1 Signaling

doi: 10.1016/j.jcmgh.2025.101629

Figure Lengend Snippet: Additional treatment with antibiotics did not affect tumor growth. ( A ) Representative macroscopic findings of the tumors, BW (g) at 20 weeks and area ratio of tumor to the whole stomach in the HFD + LPS group ( left ) and the HFD + LPS + antibiotics group ( right ) of K19-Wnt1/C2mE mice (HFD + LPS group [n = 7] vs HFD + LPS + Abx group [n = 5], bar = 5 mm). The data are represented in box plots (x = mean). Statistical analysis was performed via Student’s t- test. ( B ) Western blot analysis of the TNF-α, TLR4, and PD-L1 proteins in the tumors of the control (Ctrl), HFD + LPS, and HFD + LPS + Abx group. The results represent 3 independent experiments. ( C ) The value of the density ratio of each band to GAPDH was calculated via Image Lab (Bio-Rad), and the data are shown in box plots (x = mean). ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. ( D ) Representative images of H&E staining ( upper ; bar = 100 μm) and ZO-1 immunostaining ( lower ;, bar = 50 μm) of the ileal mucosa of the HFD + LPS group and HFD + LPS + Abx group. Mucosal atrophy with irregular villous structures in the ileum was observed following HFD + LPS administration, regardless of antibiotic treatment. ( E ) Western blot analyses of ZO-1 protein expression in the ileal mucosa. The results represent 3 independent experiments. The value of the density ratio of each band to that of β-actin was calculated via Image Lab, and the data are shown in box plots in the lower panel (x = mean). ∗∗ P < .01. Statistical analysis was conducted via the Tukey‒Kramer test. ( F ) Species richness in the HFD + LPS + Abx group compared with the HFD + LPS group (HFD + LPS group [n = 5] vs HFD + LPS + Abx group [n = 5]). The data are represented in box plots. ∗∗ P < .01. Statistical analysis was performed via the Wilcoxon rank-sum test. ( G ) PCoA plots based on weighted UniFrac distances ( left ). Weighted UniFrac distances were significantly different between the HFD + LPS + Abx group and the HFD + LPS group ( right ; HFD + LPS group [n = 5] vs HFD + LPS + Abx group [n = 5]). The values are represented in box plots. ∗∗ P < .01. Statistical analysis was performed via pairwise PERMANOVA. ( H ) Relative abundance of microbial taxa at class level in the HFD + LPS group and HFD + LPS + Abx group. The classes of ASVs not reaching 1% abundance within a sample are categorized into the “Others” group. Abx, antibiotics; Ctrl, control.

Article Snippet: Cd274/CD274 , Applied Biosystems , Mm03048248_m1, Hs00204257_m1.

Techniques: Western Blot, Control, Staining, Immunostaining, Expressing