Journal: Journal for Immunotherapy of Cancer

Article Title: AGPAT3 reshapes tumor cell vulnerability to IFNγ-mediated ferroptosis and enhances immunotherapy efficacy through lipid remodeling

doi: 10.1136/jitc-2025-013305

Figure Lengend Snippet: IFN-γ alters the ether lipid metabolism via the IRF1-AGPAT3 axis. ( A ) Schematic summarizing the PUFA-ePLs biosynthesis pathway and their contribution to ferroptosis susceptibility. ( B ) Heatmap of different groups on the expression of specific genes in A375 cells and H1299 cells. ( C ) Representative western blot for AGPS, AGPAT3 and GNPAT in A375 cells and H1299 cells after IFN-γ incubation, IFN-γ: 60 mg/mL. ( D ) Cell viability of different groups treated with different concentrations of RSL3/RSL3+Fer-1 for 24 hours after IFN-γ incubation for 48 hours (n=3), IFN-γ: 100 mg/mL, Fer-1: 1 µM. ( E ) Relative ROS level of different groups treated with RSL3/RSL3+Fer-1 for 3 hours after IFN-γ incubation for 48 hours (n=3), IFN-γ: 60 mg/mL, RSL3: 500 nM, Fer-1: 1 µM. ( F ) Heatmap of different groups on the expression of IFN-γ downstream transcription factors in A375 cells and H1299 cells. ( G–H ) IRF1 binding sites at the AGPAT3 promoter region in lung cancer cell lines from GSE186168 ChIP-seq data. ( I–J ) The peak of IRF1 binding at the Agpat3 promoter region becomes higher after IFN-γ incubation from GSE201881 ( I ) and GSE141606 ( J ) ChIP-seq data. ( K ) IRF1 binding sites at the AGPAT3 promoter region in A375 from CUT&Tag data. ChIP-seq, chromatin Immunoprecipitation sequencing; CUT&Tag, Cleavage Under Targets and Tagmentation; IFN, interferon; PUFA-ePLs, polyunsaturated ether phospholipids; ROS, reactive oxygen species.

Article Snippet: The following antibodies were used: β-actin (1:5000, Proteintech, #66009-1-Ig, China), AGPS (1:1000, Abcam, #TA321589S, USA), AGPAT3 (1:1000, Proteintech, #25723-1-AP, China), GNPAT (1:1000, #ab184186, #14931-1-AP, China), IRF1 (1:1000, CST, #8478, USA), FSP1 (1:400, Proteintech, #20886-1-AP, China), and GPX4 (1:200, Abcam, #ab125066, USA).

Techniques: Expressing, Western Blot, Incubation, Binding Assay, ChIP-sequencing

Journal: Journal for Immunotherapy of Cancer

Article Title: AGPAT3 reshapes tumor cell vulnerability to IFNγ-mediated ferroptosis and enhances immunotherapy efficacy through lipid remodeling

doi: 10.1136/jitc-2025-013305

Figure Lengend Snippet: IFN-γ alters the ether lipid metabolism via the IRF1-AGPAT3 axis. ( A–B ) Changes in ratio of ether lipids and eater lipids in A375 and H1299 cells in NC, sh-AGPAT3 and sh-AGPAT3+IFN-γ group, IFN-γ: 60 mg/mL. ( C–D ) Changes in ratio of different ether lipids in A375 and H1299 cells in NC, sh-AGPAT3 and sh-AGPAT3+IFN-γ group, IFN-γ: 60 mg/mL. ( E–F ) Heatmap of ether-PC and ether-PE in A375 ( E ) and H1299 cells ( F ) in different groups, IFN-γ: 60 mg/mL. IFN, interferon; NC, normal control; PC, phosphatidylcholine; PE, phosphatidylethanolamine.

Article Snippet: The following antibodies were used: β-actin (1:5000, Proteintech, #66009-1-Ig, China), AGPS (1:1000, Abcam, #TA321589S, USA), AGPAT3 (1:1000, Proteintech, #25723-1-AP, China), GNPAT (1:1000, #ab184186, #14931-1-AP, China), IRF1 (1:1000, CST, #8478, USA), FSP1 (1:400, Proteintech, #20886-1-AP, China), and GPX4 (1:200, Abcam, #ab125066, USA).

Techniques: Control

Journal: Journal for Immunotherapy of Cancer

Article Title: AGPAT3 reshapes tumor cell vulnerability to IFNγ-mediated ferroptosis and enhances immunotherapy efficacy through lipid remodeling

doi: 10.1136/jitc-2025-013305

Figure Lengend Snippet: Tumor AGPAT3 affects the efficacy of ICI. ( A ) Schematic diagram of in vivo experiments for IFN-γ enhancing anti-PD-1 response in mouse tumor models. ( B–C ) Average tumor growth kinetics ( B ) and tumor weights ( C ) of LLC and B16 in C1-C5 groups under different treatments (n=5). ( D–E ) The proportion of CD8 + IFN-γ + cells and CD8 + TIM3 + in T cells from LLC tumor. ( F–G ) The proportion of CD8 + IFN-γ + cells ( F ) and CD8 + TIM3 + ( G ) in T cells from B16 tumor. ( H–I ) Relative infiltration of M1 macrophages and M2 macrophages and M1/M2 ratio in the TIME of each group from LLC tumor ( H ) (n=5) and B16 tumor ( I ) (n=5). FACS, fluorescenceactivated cell sorting; IFN, interferon; LLC, lewis lung carcinoma; PBS, phosphate-buffered saline; PD-1, programmed cell death protein-1; TIME, tumor immune microenvironment; .

Article Snippet: The following antibodies were used: β-actin (1:5000, Proteintech, #66009-1-Ig, China), AGPS (1:1000, Abcam, #TA321589S, USA), AGPAT3 (1:1000, Proteintech, #25723-1-AP, China), GNPAT (1:1000, #ab184186, #14931-1-AP, China), IRF1 (1:1000, CST, #8478, USA), FSP1 (1:400, Proteintech, #20886-1-AP, China), and GPX4 (1:200, Abcam, #ab125066, USA).

Techniques: In Vivo, FACS, Saline

Journal: Journal for Immunotherapy of Cancer

Article Title: AGPAT3 reshapes tumor cell vulnerability to IFNγ-mediated ferroptosis and enhances immunotherapy efficacy through lipid remodeling

doi: 10.1136/jitc-2025-013305

Figure Lengend Snippet: The clinical and immunological roles of AGPAT3 across cancer types. ( A ) Correlation between AGPAT3 expression and FD.score in TCGA pan-cancer. ( B ) Correlation between AGPAT3 expression and ferroptosis-related gene in TCGA-LUSC and TCGA-SKCM. ( C ) Correlation between AGPAT3 expression and CD8 + Teff score in HNSC, LUSC and UVM of TCGA database. ( D ) Correlation between AGPAT3 expression and IFNG in ICI cohorts. ( E ) Survival analysis of AGPAT3 expression in the Snyder2017 cohort. ( F ) Comparison of AGPAT3 expression between responder and non-responder in GSE165278 and IMvigor210. ESCA, esophageal carcinoma; FD.score, ferroptosis-driver signature score; GBM, glioblastoma; HNSC, head and neck squamous carcinoma; ICI, immune checkpoint inhibitor; IFN, interferon; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LGG, lower grade glioma; LIHC, liver hepatocellular carcinoma; LUSC, lung squamous cell carcinoma; NR, non-responders; OV, ovarian serous cystadenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD, prostate adenocarcinoma; R, responders; SARC, sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TCGA, The Cancer Genome Atlas; TGCT, testicular germ cell tumors; THCA, thyroid carcinoma; UCEC, uterine corpus endometrial carcinoma; UVM, uveal melanoma.

Article Snippet: The following antibodies were used: β-actin (1:5000, Proteintech, #66009-1-Ig, China), AGPS (1:1000, Abcam, #TA321589S, USA), AGPAT3 (1:1000, Proteintech, #25723-1-AP, China), GNPAT (1:1000, #ab184186, #14931-1-AP, China), IRF1 (1:1000, CST, #8478, USA), FSP1 (1:400, Proteintech, #20886-1-AP, China), and GPX4 (1:200, Abcam, #ab125066, USA).

Techniques: Expressing, Comparison

Journal: Translational Gastroenterology and Hepatology

Article Title: ORM2 protects against acute pancreatitis by inhibiting premature activation of pancreatic enzymes

doi: 10.21037/tgh-25-106

Figure Lengend Snippet: Reduced pancreatic ORM2 expression in AP. (A) Scatterplots showing log2 transformation of the normalized blood ORM1 in patients with MAP (n=57) and healthy controls (n=32). (B) Scatterplots showing log2 transformation of the normalized blood ORM2 in patients with MAP (n=57) and healthy controls (n=32). (C) The serum ORM in control and AP mice (n=6 per group). (D) Immunoblotting of ORM1 and ORM2 in serum from control and AP mice (n=6 per group). (E) Immunoblotting and quantitative analysis of ORM1 and ORM2 in pancreas from control and AP mice (n=6 per group). (F) Immunoblotting and quantitative analysis of ORM1 and ORM2 in liver from control and AP mice (n=6 per group). Data are presented as mean ± SEM. The groups were compared using unpaired two-tailed student’s t -test (*, P<0.05; ***, P<0.001; ****, P<0.0001). AP, acute pancreatitis; MAP, mild acute pancreatitis; ORM, orosomucoid; SEM, standard error of the mean.

Article Snippet: The antibodies used in this study are listed as follows: ORM1 (No. 16439-1-AP proteintech, Wuhan, China), ORM2 (No. P19652 , zenbio, Chengdu, China), GAPDH (No. A19056, ABclonal, Wuhan, China), HRP-conjugated Goat anti-Rabbit IgG (H+L) (No. AS014, ABclonal, Wuhan, China).

Techniques: Expressing, Transformation Assay, Control, Western Blot, Two Tailed Test

Journal: Molecular Biology of the Cell

Article Title: Interplay of septin amphipathic helices in sensing membrane-curvature and filament bundling

doi: 10.1091/mbc.E20-05-0303

Figure Lengend Snippet: Genetic analyses of Shs1 and Cdc12 AH domains. (A) Viability of cdc12-6 cells expressing indicated SHS1 alleles expressed from the endogenous locus (with 3xHA tag, unless indicated with GFP tag, which lacks 3xHA) based on tetrad dissections at permissive temperature (24°C). Cells from genotypes labeled in blue appeared normal, without obvious septin defects. Genotypes in orange were sick, with partially or fully penetrant septin defects. Genotypes labeled in red were inviable. (B) DIC images of cdc12-6 shs1 mutants (see A) at 24°C. Scale bar, 5 μm. (C) Heterozygous diploids expressing the indicated Shs1 protein fused to GFP from the SHS1 locus. Scale bar, 5 μm. (D) Scatter plot quantifying cdc12-6-SpoVM septin complex adsorption onto different membrane curvatures. Black bars represent the mean. Error bars are the SD for more than 30 measured beads at each curvature across three replicates. Adsorption of cdc12-6-SpoVM complexes was significantly greater on a membrane curvature of 2 μm −1 than on curvatures of 0.67 and 0.4 μm −1 ; ** ( p < 0.01 and p < 0.0001, respectively). ns, adsorption was not significantly different. (E) Western blot comparing expression of indicated Cdc12 chimeras fused to 3xHA epitope in heterozygous diploids.

Article Snippet: A polyclonal rabbit α-HA epitope antibody preparation was used at 1:2000 dilution (Rockland Immunochemicals).

Techniques: Expressing, Labeling, Adsorption, Membrane, Western Blot

Journal: Neuroscience Bulletin

Article Title: Comprehensive Proteomic Profiling of Patients’ Tears Identifies Potential Biomarkers for the Traumatic Vegetative State

doi: 10.1007/s12264-018-0259-x

Figure Lengend Snippet: Gene Ontology analysis revealed 21 proteins involved in response to wounding.

Article Snippet: In the verification stage, the levels of 7 promising tear proteins [cystatin B (CTSB), protease, serine 1 (PRSS1), S100 calcium-binding protein A7 (S100A7), glutathione S-transferase P (GSTP1), complement factor H (CFH), kininogen 1 (KNG1), and alpha-1-acid glycoprotein 1 (ORM1)] were measured using the ELISA kits for human CSTB, human PRSS1, human CFH, human KNG1, and human ORM1 (Boster Biological Technology, Wuhan, China) and for human S100A7 and human GSTP1 (from Cloud-clone Corp., Houston, TX).

Techniques:

Journal: Neuroscience Bulletin

Article Title: Comprehensive Proteomic Profiling of Patients’ Tears Identifies Potential Biomarkers for the Traumatic Vegetative State

doi: 10.1007/s12264-018-0259-x

Figure Lengend Snippet: Levels of representative proteins in tears from healthy controls and traumatic vegetative state patients. A List of 7 selected differentially-expressed proteins. Levels of CTSB (B), PRSS1 (C), S100A7 (D), GSTP1 (E), CFH (F), KNG1 (G), and ORM1 (H).

Article Snippet: In the verification stage, the levels of 7 promising tear proteins [cystatin B (CTSB), protease, serine 1 (PRSS1), S100 calcium-binding protein A7 (S100A7), glutathione S-transferase P (GSTP1), complement factor H (CFH), kininogen 1 (KNG1), and alpha-1-acid glycoprotein 1 (ORM1)] were measured using the ELISA kits for human CSTB, human PRSS1, human CFH, human KNG1, and human ORM1 (Boster Biological Technology, Wuhan, China) and for human S100A7 and human GSTP1 (from Cloud-clone Corp., Houston, TX).

Techniques: