Immunohistochemical studies on surgically resected gliomas (Gliomas 1, 5, 9; Table 1) and non-diseased human frontal lobe (n = 3), revealing GPIHBP1 expression in capillaries of gliomas but not in …
Immunohistochemical studies on glioma sample 5 revealed GPIHBP1 in capillaries (detected by mAb RF4; left). In the control panel (right), the section was handled in the same fashion, except that the …
(A) Confocal fluorescence microscopy studies on sections from glioma sample 1 (Table 1), demonstrating the detection of GPIHBP1 with three different human GPIHBP1–specific monoclonal antibodies …
Confocal microscopy images of a BFP-tagged CT-2A glioma implanted in a ROSAmT/mG::Pdgfb-iCreERT2 mouse, revealing the expression of GPIHBP1 in capillary endothelial cells of the glioma but not those …
Confocal immunofluorescence imaging of a BFP-tagged CT-2A glioma and adjacent normal brain in a ROSAmT/mG::Pdgfb-iCreERT2 mouse, revealing GPIHBP1 in the capillary endothelial cells of the glioma …
A four-month-old C57BL/6 mouse harboring a CT-2A glioma was euthanized and perfusion-fixed with carbodiimide/glutaraldehyde before sections of glioma and brain were processed for TEM. Electron …
The endothelial cells were cultured in medium in the presence or absence of mouse VEGF (100 ng/ml) for 24 hr. (A) Transcript levels for Gpihbp1, Cd31 (an endothelial cell gene), and three genes …
Immunohistochemical studies of a BFP-expressing CT-2A glioma (after three weeks of growth). Mice were injected via the tail vein with an Alexa Fluor 647–labeled antibody against mouse GPIHBP1 (11A12;…
Immunohistochemical studies on a CT-2A glioma collected from a mouse that had been given an intravenous injection of a GPIHBP1-specific antibody. Tissue sections (200-μm-thick) were fixed with 2% …
Immunohistochemical studies on mouse glioma and normal brain, revealing GPIHBP1 in gliomas but not normal brain. Tissue sections (10-μm-thick) were fixed with 3% PFA in PBS and stained with …
RNA expression data for normal mouse brain vascular cells and mouse glioma vascular cells reveal high levels of Gpihbp1 expression in the endothelial cells of gliomas but not those of normal brain. G…
Immunohistochemical studies of BFP-expressing CT-2A gliomas (after three weeks of growth) in Gpihbp1+/+ and Gpihbp1–/– mice. Mice were perfused with PBS and perfusion-fixed with 2% PFA. Glioma and …
Confocal immunofluorescence microscopy studies on glioma and normal brain from wild-type and Gpihbp1–/– mice, along with the brain from an Lpl–/– mouse carrying a skeletal muscle–specific human LPL …
Confocal immunofluorescence microscopy studies on a mouse glioma (10-μm section), revealing large numbers of macrophages (detected with an antibody against F4/80 [red]) in the glioma. Endothelial …
Peritoneal macrophages were harvested and plated on poly-D-lysine–coated glass coverslips and then fixed with 3% PFA. Using a goat antibody against mouse LPL, LPL (red) was detected in macrophages …
Brain and glioma from wild-type mice and brain from Lpl–/–MCK-hLPL (MCK) mice were harvested, sectioned (10-μm), and fixed with 3% PFA. LPL (detected with a goat antibody against mouse LPL [red]) …
Rows in the heat map correspond to individual genes; columns correspond to individual specimens, normal brain specimens on the left (GTEx; green) and tumors on the right (TCGA; purple). LPL was the …
Sections (10-μm) of brain and glioma tissue from wild-type and Gpihbp1–/– mice and brain tissue from Lpl–/–MCK-hLPL mice (MCK) were fixed with 3% PFA and stained with a goat antibody against mouse …
Sections (10-μm) of heart and brain from Lpl–/–MCK-hLPL mice and of heart tissue from wild-type mice were fixed with 3% PFA and stained with a goat antibody against mouse LPL, a mAb against GPIHBP1 …
Sections (10-μm) of brain specimens from wild-type and Lpl–/–MCK-hLPL mice were fixed with 3% PFA and stained with antibodies against LPL (red) and CD31 (green). The left panel shows a tiled …
Mouse Gpihbp1 (A), mouse Lpl (B), and human LPL (C) transcript levels, as measured by qRT-PCR (n = 3/group). Expression levels were normalized to the expression of cyclophilin A. BAT, brown adipose …
Four-month-old C57BL/6 mice harboring CT-2A gliomas were fasted for 4 hr and then injected intravenously with 200 μl of [2H]TRLs. After 1 min, mice were euthanized and perfusion-fixed with …
Four-month-old C57BL/6 mice harboring CT-2A gliomas were fasted for 4 hr and then injected intravenously with 200 μl of [2H]TRLs. After 30 min, mice were euthanized and perfusion-fixed with …
Four-month-old C57BL/6 mice harboring CT-2A gliomas were fasted for 4 hr and then injected intravenously with 200 μl of PBS. After 1 min, the mouse was euthanized and perfusion-fixed with …
A C57BL/6 wild-type mouse (Gpihbp1+/+) and a Gpihbp1-deficient mouse (Gpihbp1–/–) were injected intravenously with 80 μl of [2H]TRLs. After 15 min, the mouse was euthanized; perfusion-fixed with …
(A) NanoSIMS images showing 13C enrichment in mouse tissues (brain, glioma, and heart) after oral administration of 13C-labeled mixed fatty acids to mice (three 80-mg doses administered 12 h apart). …
13C-labeled mixed fatty acids were administered by gastric gavage to C57BL/6 mice harboring CT-2A gliomas (three doses of 80 mg administered 12 hr apart). Four hours after the last dose, the mice …
Four hours after the last dose, mice were euthanized; perfusion-fixed with carbodiimide/glutaraldehyde; and sections of normal brain were prepared for NanoSIMS imaging. 12C14N– images were generated …
Four-month-old C57BL/6 mice harboring CT-2A gliomas were fasted for 4 hr and then injected intravenously with 200 μl of PBS. After 1 min, the mouse was euthanized; perfusion-fixed with …
CT-2A glioma cells that were stably transfected with a Gaussia luciferase reporter were injected intracranially into Gpihbp1+/+ (blue) and Gpihbp1–/– (red) mice (n = 11/group). Tumor growth in live …
In normal brain (left panel), LPL is produced by astrocytes, neurons, oligodendrocytes, and fibroblasts. Because GPIHBP1 is not expressed in the capillaries of the brain parenchyma, we have proposed …
Expression of GPIHBP1 was assessed by immunohistochemistry with mAbs against human GPIHBP1 (RF4, RE3, RG3). Those conducting the studies were blinded to diagnoses. This table details the tumor …
Sample ID | Tissue diagnosis | Location | 1p/19q co-deletion | IDH1 mutation | GPIHBP1 |
---|---|---|---|---|---|
1 | Glioblastoma (GBM) | Right frontal, parietal | No | Negative | Yes |
2 | GBM | Left temporal | No | Negative | Yes |
3 | GBM | Right occipital | No | Negative | Yes |
4 | GBM | Left frontal | No | Negative | Yes |
5 | Oligodendroglioma Grade II | Left anterior temporal, left posterior temporal | Yes | Negative | Yes |
6 | Oligoastrocytoma Grade III | Right temporal | No | Negative | Yes |
7 | GBM + oligodendroglial component | Left frontal | Yes | Negative | Yes |
8 | GBM + extensive oligodendroglial component | Right frontal | No | Negative | Yes |
9 | Oligodendroglioma Grade III | Left frontal | Yes | +R132H | Yes |
10 | Oligodendroglioma Grade III | Left frontal | Yes | +R132H | Yes |
11 | Oligoastrocytoma | Right parietal | No | Negative | Yes |
12 | Oligodendroglioma Grade III | Right parietal | Yes | +R132H | Yes |
13 | Oligodendroglioma Grade III | Right parietal | Yes | Negative | Yes |
14 | Oligoastrocytoma Grade III | Left temporal | No | +R132H | Yes |
15 | Oligoastrocytoma Grade III | Right temporal | No | +R132G | No |
16 | Oligoastrocytoma Grade III | Right frontal | No | +R132H | No |
17 | Oligodendroglioma Grade III | Left frontal | Yes | Negative | No |
18 | Oligodendroglioma Grade III | Left frontal | Yes | +R132H | No |
19 | Oligodendroglioma Grade III | Left temporal | Yes | Negative | No |
20 | Oligodendroglioma Grade III | Right temporal | Yes | +R132H | No |
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (M. musculus) | Gpihbp1–/– | Beigneux et al., 2007 | RRID: MGI:3771172 | Dr. Stephen G Young (UCLA) |
Genetic reagent (M. musculus) | Lpl–/–MCK-hLPL | Levak-Frank et al., 1995 | RRID: MGI:3624988 | Dr. Rudolph Zechner (Graz University) |
Genetic reagent (M. musculus) | ROSAmT/mGPdgfb-iCreT2 | Mathivet et al., 2017 | Dr. Holger Gerhardt (VIB KU-Leuven) | |
Cell line (M. musculus) | CT-2A | Seyfried et al., 1992 | Dr. Thomas Seyfried (Boston College) | |
Cell line (M. musculus) | CT-2A–BFP | PMID: 24658686 | Dr. Holger Gerhardt (VIB KU-Leuven) | |
Cell line (M. musculus) | bEnd.3 | ATCC | Catalog No. CRL-2299 RRID: CVCL_0170 | |
Transfected construct (lentiviral plasmid) | plenti-GLuc-IRES-EGFP | Targeting Systems | Catalog No. GL-GFP | |
Antibody | Rat monoclonal anti-mouse GPIHBP1 (11A12) | Beigneux et al., 2009 | Dr. Stephen G Young (UCLA); IHC (10 μg/ml) | |
Antibody | Mouse monoclonal anti-human GPIHBP1 (RE3) | Hu et al., 2017 | Dr. Stephen G Young (UCLA); IHC (10 μg/ml) | |
Antibody | Mouse monoclonal anti-human GPIHBP1 (RF4) | Hu et al., 2017 | Dr. Stephen G Young (UCLA); IHC (10 μg/ml) | |
Antibody | Mouse monoclonal anti-human GPIHBP1 (RG3) | Hu et al., 2017 | Dr. Stephen G Young (UCLA); IHC (10 μg/ml) | |
Antibody | Rabbit polyclonal anti-vWF | Dako | Catalog No. A0082 RRID: AB_2315602 | IHC (1:200) |
Antibody | Goat polyclonal anti-GFAP | Abcam | Catalog No. ab53554 RRID: AB_880202 | IHC (1:200) |
Antibody | Rabbit polyclonal anti-GLUT1 | Millipore-Sigma | Catalog No. 07–1401 RRID: AB_1587074 | IHC (1:200) |
Antibody | Rabbit polyclonal anti-CD31 | Abcam | Catalog No. ab28364 RRID: AB_726362 | IHC (1:50) |
Antibody | Rat monoclonal anti-F4/80 | Abcam | Catalog No. ab6640 RRID: AB_1140040 | IHC (10 μg/ml) |
Antibody | Goat polyclonal anti-mouse LPL | Page et al., 2006 | Dr. André Bensadoun (Cornell); IHC (12 μg/ml) | |
Antibody | Alexa Fluor 488, 568, 647 secondaries | ThermoFisher Scientific | IHC (1:500) | |
Commercial assay or kit | ImmPRESS Excel Staining Kit | Vector Laboratory | Catalog No. MP-7602 | |
Sequence-based reagent | Mouse Gpihbp1 primers | 5′-AGCAGGGACAGAGCACCTCT-3′ and 5′-AGACGAGCGTGATGCAGAAG-3′ | ||
Sequence-based reagent | Mouse Cd31 primers | 5′-AACCGTATCTCCAAAGCCAGT-3′ and 5′-CCAGACGACTGGAGGAGAACT-3′ | ||
Sequence-based reagent | Mouse Angpt2 primers | 5′-AACTCGCTCCTTCAGAAGCAGC-3′ and 5′-TTCCGCACAGTCTCTGAAGGTG-3′ | ||
Sequence-based reagent | Mouse Dusp5 primers | 5′-TCGCCTACAGACCAGCCTATGA-3′ and 5′-TGATGTGCAGGTTGGCGAGGAA-3′ | ||
Sequence-based reagent | Mouse Cxcr4 primers | 5′-GACTGGCATAGTCGGCAATGGA-3′ and 5′-CAAAGAGGAGGTCAGCCACTGA-3′ | ||
Sequence-based reagent | Mouse Lpl primers | 5′-AGGTGGACATCGGAGAACTG-3′ and 5′-TCCCTAGCACAGAAGATGACC-3′ | ||
Sequence-based reagent | Human LPL primers | 5′-TAGCTGGTCAGACTGGTGGA-3′ and 5′-TTCACAAATACCGCAGGTG-3′ | ||
Recombinant DNA reagent | ALO-D4 plasmid | Gay et al., 2015 | Dr. Arun Radhakrishnan (UT Southwestern) | |
Chemical compound, drug | N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (carbodiimide) | Millipore-Sigma | Catalog No. 03449 | |
Chemical compound, drug | Glutaraldehyde25% solution | Electron Microscopy Sciences | Catalog No. 16220 | |
Chemical compound, drug | Osmium tetroxide 4% solution | Electron Microscopy Sciences | Catalog No. 18459 | |
Chemical compound, drug | Paraformaldehyde 16% solution | Electron Microscopy Sciences | Catalog No. 15170 | |
Chemical compound, drug | EMbed 812 | Electron Microscopy Sciences | Catalog No. 14120 | |
Chemical compound, drug | Sodium cacodylate trihydrate | Electron Microscopy Sciences | Catalog No. 12300 | |
Chemical compound, drug | Uranyl acetate | SPI-Chem | Catalog No. 02624AB | |
Chemical compound, drug | DAPI | ThermoFisher Scientific | Catalog No. 1306 | IHC (3 μg/ml) |
Chemical compound, drug | Mouse VEGF | Millipore-Sigma | Catalog No. V4512 | |
Software, algorithm | LIMMA | Ritchie et al., 2015 | RRID: SCR_010943 | |
Other | D-GLUCOSE (U-13C6, 99%) | Cambridge Isotope Laboratories | Catalog No. CLM-1396-PK | |
Other | Mixed fatty acids (U-D, 96–98%) | Cambridge Isotope Laboratories | Catalog No. DLM-8572-PK | |
Other | Mixed fatty acids (13C, 98%+) | Cambridge Isotope Laboratories | Catalog No. CLM-8455-PK |