Journal: Journal of Lipid Research

Article Title: Peroxisomal ether-glycerophospholipid synthesis is dysregulated after TBI

doi: 10.1016/j.jlr.2025.100821

Figure Lengend Snippet: Peroxisomal ether-GPs synthesis was disrupted after TBI. A: schematic diagram showing peroxisomal ether-GPs synthesizing steps. It is initiated by the acylation of dihydroxyacetone phosphate (DHAP) by GNPAT that generates 1-acyl-DHAP which is then converted to 1-O-alkyl-DHAP by AGPS. 1-O-alkyl-DHAP is then converted to 1-O-alkylglycerol phosphate which is transported to the endoplasmic reticulum for the generation of fully formed ether-GPs. B: Western blots of peroxisomal enzymes GNPAT and AGPS in the cortical tissue lysates of sham and TBI mice. Data presented as mean ± SEM. n = 4; ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. C: Western blots and (D) corresponding quantification of peroxisomal enzymes GNPAT and AGPS in the peroxisomal and cytosolic fractions prepared from the sham and TBI (PID 1) mouse cortices. ABCD3 (PMP70) and PEX14 are markers of peroxisomal membrane and α-tubulin is of cytosolic fraction. Data presented as mean ± SEM. n = 4. ∗∗ P < 0.01 and ∗ P < 0.05 with respect to sham determined by One-way ANOVA. E: Western blots and corresponding (F) quantification of GNPAT and AGPS in the peroxisomal (ABCD3+) and cytosolic (α-tubulin+) fractions of sham and TBI mouse cortices (PID 28). Data = Mean ± SEM; n = 5. ∗∗ P < 0.01, Students' t test. G: 60X images of AGPS and peroxisomal membrane marker PEX14. H: Quantification of punctate versus diffused AGPS ratio in sham (blue) and injured (1 day after TBI) (red) brain sections. Data presented as mean ± SEM. n = 3; ∗ P < 0.05; Students' t test.

Article Snippet: Primary antibodies: AGPS (1:1000; Thermo Scientific, PA5-87935), GNPAT (1:1000; Thermo Scientific, PA5-36447), ABCD3/PMP70 (1:1000, Thermo Scientific, PA1-650), α-Tubulin (1:500; AA4.3-s, developed by Walsh, C. and obtained from Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biology, Iowa City, IA 52,242), β-actin/ACTB (1:10,000; Sigma, A1978) and PEX7 (1:1,000, Proteintech, 20614-1-AP) and PEX14 (1:1000, Proteintech, 10594-1-AP).

Techniques: Western Blot, Membrane, Marker

Journal: Cancer Science

Article Title: Abnormal Vessels Potentially Accelerate Glioblastoma Proliferation by Inducing the Protumor Activation of Macrophages

doi: 10.1111/cas.70014

Figure Lengend Snippet: Mouse experiments using glioma cell lines and spatial analysis of human GBM. (A) Schema of mouse experiment. (B) Survival curve of WT and AGP KO mice after intracranial injection of GL261. (C) CD34 immunostaining of brain tissue from WT and AGP KO mice in the GL261 injection group and control group. (D) Comparison of tumor vascular area between WT and AGP KO mice in the GL261 injection group.

Article Snippet: Plasma AGP of GBM patients and healthy donors were detected by sandwich ELISA using an anti‐ORM1 antibody (16439‐1‐AP; Proteintech, IL, USA) and labeled with Biotin Labeling Kit—NH 2 (Dojindo).

Techniques: Injection, Immunostaining, Control, Comparison

Journal: Cancer Science

Article Title: Abnormal Vessels Potentially Accelerate Glioblastoma Proliferation by Inducing the Protumor Activation of Macrophages

doi: 10.1111/cas.70014

Figure Lengend Snippet: Expression of PLVAP and AGP in relation to GBM malignancy. (A) Double immunostaining and immunofluorescence showing the expression of Ki67 around PLVAP‐positive vessels. The lower panel shows a comparison of Ki67 positivity around PLVAP‐positive and negative vessels. (B) IHC of PLVAP in normal brain and glioma grades 2–4. (C) Plasma AGP levels in normal adults and human GBM patients. (D) IHC of AGP in normal brain and glioma grades 2–4. (E) AGP leakage around vessels in untreated GBM cases. (F) Comparison of AGP immunostaining before and after Bev treatment. ** p < 0.01, **** p < 0.0001.

Article Snippet: Plasma AGP of GBM patients and healthy donors were detected by sandwich ELISA using an anti‐ORM1 antibody (16439‐1‐AP; Proteintech, IL, USA) and labeled with Biotin Labeling Kit—NH 2 (Dojindo).

Techniques: Expressing, Double Immunostaining, Immunofluorescence, Comparison, Clinical Proteomics, Immunostaining

Journal: Cancer Science

Article Title: Abnormal Vessels Potentially Accelerate Glioblastoma Proliferation by Inducing the Protumor Activation of Macrophages

doi: 10.1111/cas.70014

Figure Lengend Snippet: Schematic representation of the findings from this study. In GBM, before BBB disruption occurs, PLVAP in the vascular endothelium is negative, Claudin‐5 is positive, and AGP is present only within the blood vessels. Tumor angiogenesis is promoted by VEGF released from tumor cells and GAMs. However, once BBB disruption occurs, the vascular endothelium undergoes abnormal proliferation, PLVAP becomes positive, and Claudin‐5 becomes negative. PLVAP becomes positive. AGP leaks into the tumor stroma, activating GAMs and promoting tumor growth. Bevacizumab normalizes blood vessels, leading to PLVAP negativity and a reduction in GAM activity.

Article Snippet: Plasma AGP of GBM patients and healthy donors were detected by sandwich ELISA using an anti‐ORM1 antibody (16439‐1‐AP; Proteintech, IL, USA) and labeled with Biotin Labeling Kit—NH 2 (Dojindo).

Techniques: Disruption, Activity Assay