Journal: bioRxiv

Article Title: TMEM Doorway Mediated Metastasis in Pancreatic Ductal Adenocarcinoma by Tie2 Signaling

doi: 10.64898/2025.12.05.692608

Figure Lengend Snippet: (A) Immunofluorescence imaging of Tie2Hi/VEGFAHi macrophages in TMEM doorways in sequential sections. Endothelial cells (EC), macrophage (M) and tumor tissue (TC). In the merge panel, a mask for IBA1 macrophage was created in ImageJ ® for a better visualization of Tie2. Scale bar,10μm. (B and C) Quantification of Tie2+ macrophages in tumor area and withim TMEM doorway area of KPCY T-low (B) and KPC (C) tumors. -total macrophages. -Tie2 positive macrophages. (D) Western blot analysis of phospho-Tie2 immunoprecipitated from extracts of KPC, KPCY T-low, RAW 264.7 or 3B-11 cells treated with Rebastinib +/- Angiopoietin 2. (E) Experimental design of the subluminal to luminal trans-endothelial migration (iTEM) assay. Macrophages = red. Tumor cells = green. The tumor cells (green) are considered transendothelial migrating cells when they are crossing the endothelial layer (pink). (F) Representative image of iTEM assay from confocal microscope. Macrophages = red. Tumor cells = green. Endothelial layer = pink. Quantitation of trans-endothelial migration of KPCY T-low (G) and KPC (H). In KPCY T-low and KPC, we observed an increase of transendothelial migration from 3 cells/field to 12.65 cells/field and 6 cells/field to 16.65 cells/field respectively, in the presence of macrophages (p<0.05), following by a decrease to 8.46 cells/field and 8.46 cells/field after rebastinib treatment (p<0.05). Co-culture with macrophages significantly increased migration, which was inhibited by the Tie2 inhibitor rebastinib. Data are presented as mean ± SEM; (n ≥ 10 per group). *P < 0.05, by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: KPC ( Cat.#: 153600: Cancer tools.org ) , KPCY T-low (6419c5) , or KPCY C-EMT 3077 (provided by Ben Z. Stanger) , murine macrophage cell line RAW264.7 (ATCC-provided by Dianne Cox), and murine endothelial cells 3B11 (ATCC - provided by Camille Duran) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (cat# SH30243, Hyclone, GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS) (cat# S11550, Atlanta Biologicals, Flowery Branch, GA, USA).

Techniques: Immunofluorescence, Imaging, Western Blot, Immunoprecipitation, Migration, Microscopy, Quantitation Assay, Co-Culture Assay

Journal: bioRxiv

Article Title: TMEM Doorway Mediated Metastasis in Pancreatic Ductal Adenocarcinoma by Tie2 Signaling

doi: 10.64898/2025.12.05.692608

Figure Lengend Snippet: (A) panel 1 shows TMEM doorways, visualized by immunohistochemistry (IHC) staining for Mena, Iba-1, and endomucin. The three cells of the TMEM doorway (contained in black circle, with the three cells forming the TMEM doorway indicated with the yellow triangle) are the TMEM doorway endothelial cell (TEC, endomucin stained in blue), TMEM doorway macrophage (TM, Iba1 stained in brown), and TMEM doorway tumor cell (TTC, Mena stained in pink). Panels 2-4 show different staining channels of the sequential tissue section 2 aligned with the IHC stained section (panel 1) with immunofluorescence (IF) with antibodies against endomucin (panel 3 yellow), dextran (panel 4 pink) and nuclear stain DAPI (blue). Active versus inactive TMEM doorways were distinguished by the presence of extravascular dextran staining (panel), which indicates that the vessel had a TMEM doorway-associated vascular opening (TAVO). Scale bars=20μm. (B-H) Mice were treated with control chow or ∼0.44 mg/day of rebastinib for 3 weeks. (B and D) Quantification of TMEM doorway density in 10 high-power fields (HPFs; no significant difference) in KPCY T-low (B) and KPC tumors (D). (C and E) Immunofluorescence measurement of extravascular dextran levels in Tie2 + TMEM doorways indicate significant inhibition of TMEM doorways by rebastinib inKPCY T – low tumors (C) and KPC tumors (E). TMEM doorways were identified in the IF-stained sections as described above. Active TMEM doorways were identified by the presence of extravascular dextran which is not present in inactive TMEM doorways. *p<0.05. (F) Disseminated tumor cells in the liver stained with p53 (red). Increased magnification of field of the p53 positive cells is shown in the yellow box. The left bottom is a representative image of control and right bottom of rebastinib treated group in magnified fields. (G-H) Quantification of DTCs in the liver of animals as compared to the control with rebastinib treated groups showing significant inhibition of DTC levels. (G) KPCY T-low (H) KPC where each dot is a separate mouse. *p<0.05, analyzed by Student’s t-test. N=8.

Article Snippet: KPC ( Cat.#: 153600: Cancer tools.org ) , KPCY T-low (6419c5) , or KPCY C-EMT 3077 (provided by Ben Z. Stanger) , murine macrophage cell line RAW264.7 (ATCC-provided by Dianne Cox), and murine endothelial cells 3B11 (ATCC - provided by Camille Duran) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (cat# SH30243, Hyclone, GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS) (cat# S11550, Atlanta Biologicals, Flowery Branch, GA, USA).

Techniques: Immunohistochemistry, Staining, Immunofluorescence, Control, Inhibition

Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: Low 15‐hydroxyprostaglandin dehydrogenase (15‐ PGDH ) expression is implicated in poor pancreatic ductal adenocarcinoma ( PDAC ) prognosis. A, Workflow diagram of patients who underwent pancreatic resection and contributed samples for immunohistochemical ( IHC ) analysis. B, Representative IHC staining of 15‐ PGDH expression in 107 PDAC tissues. Scale bar = 100 μm. C,D, Relationship between 15‐ PGDH expression and relapse‐free survival (C) or overall survival (D) using the Kaplan‐Meier method

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry

Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: 15‐Hydroxyprostaglandin dehydrogenase (15‐ PGDH ) downregulation by interleukin‐1β ( IL ‐1β) enhances pancreatic ductal adenocarcinoma cell growth. A,B, Expression of HPGD (the gene coding 15‐ PGDH protein, upper panel) or 15‐ PGDH (lower panel) in PK ‐8 cells (A) or S2‐013 cells (B) after treatment with si RNA targeting 15‐ PGDH or with control si RNA , evaluated by quantitative RT ‐ PCR (upper panel) or Western blot analysis (lower panel). Data are presented as the treated/control cell ratio. C,D, PK ‐8 cells (C) or S2‐013 cells (D) transfected with si RNA s targeting 15‐ PGDH or with control si RNA were incubated for up to 96 hours and assayed for cell number; data are presented as the treated/control (time = 0) cell ratio. E,F, Expression of 15‐ PGDH in PK ‐8 cells or S2‐013 cells after IL ‐1β (E) or tumor necrosis factor‐α ( TNF ‐α) (F) treatment for 24 and 48 hours and distilled water treatment for 48 hours as a control was evaluated by Western blotting. G, Column graph showing relative 15‐ PGDH levels in PK ‐8 cells or S2‐013 cells after IL ‐1β and TNF ‐α treatment for 24 and 48 hours, and distilled water treatment for 48 hours as a control, were evaluated using ImageJ software. H, Expression of HPGD and IL 1B in six PDAC patients determined by quantitative RT ‐ PCR . Data were normalized to the ACTB mRNA level and are shown as the mean ± SD of three independent experiments. **P < .01

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Transfection, Incubation, Software

Journal: Cancer Science

Article Title: Downregulation of 15‐hydroxyprostaglandin dehydrogenase by interleukin‐1β from activated macrophages leads to poor prognosis in pancreatic cancer

doi: 10.1111/cas.13467

Figure Lengend Snippet: Tumor‐associated macrophages are inversely correlated with pancreatic ductal adenocarcinoma ( PDAC ) cells harboring high 15‐hydroxyprostaglandin dehydrogenase (15‐ PGDH ) expression. A,B, Representative immunohistochemical ( IHC ) staining of 15‐ PGDH (upper panel) and CD 163 (lower panel) expression in high 15‐ PGDH (A) and low 15‐ PGDH (B) serial PDAC specimens. Scale bar = 200 μm. C, Graph showing Pearson's correlation between the expression of 15‐ PGDH and the number of CD 163‐positive cells in 107 PDAC patients. D, Schematic representation of the findings of this study. IL ‐1βR, interleukin‐1β receptor

Article Snippet: The human PDAC cell lines PK‐8 and S2‐013 were obtained from the Japanese Collection of Research Bioresource Cell Bank (Ibaraki, Japan) and RIKEN Bioresource Center Cell Bank (Tsukuba, Japan).

Techniques: Expressing, Immunohistochemical staining, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: CTC-derived pancreatic cancer models serve as research tools and are suitable for precision medicine approaches

doi: 10.1016/j.xcrm.2024.101692

Figure Lengend Snippet: Highly efficient retrieval of CTCs from patients with pancreatic cancer (A) Comparison of CTC capture rates using EpCAM antibody-coated beads and beads with additional antibodies in PDX-derived primary PDAC cultures (SiC024). EMT induction in PDAC cultures using macrophage-conditioned media (MCM) served two purposes: mimicking clinical heterogeneity and simulating metastatic conditions. During metastasis, cancer cells often downregulate EpCAM expression, rendering EpCAM-only isolation procedures less effective. To model CTC analysis, 100 PDAC cells were spiked into 5 mL of blood obtained from healthy individuals. Beads coated with varying antibody combinations (EpCAM alone, EpCAM + CD47, EpCAM + FAP, and a triple combination of EpCAM, CD47, and FAP antibodies) were employed to isolate CTCs. The graph illustrates the count of isolated spiked-in PDAC cells as a model for CTCs ( n = 5). Boxplots show median values as horizontal lines within the boxes, with whiskers representing the 95% confidence intervals (CI). (B) Comparison of CTC retrieval efficiency of 100 cultured PDAC cells (SiC024) spiked into 5 mL of healthy blood. Three methods were used: triple antibody-coated beads, VAR2-coated beads, and immunodensity cell isolation by RosetteSep. The graph shows the number of retrieved CTCs ( n = 5). (C) Analysis of CTC retrieval from four patients with PDAC. The same volume of patient-derived blood (5 mL) was used for each isolation. Beads coated with EpCAM or triple antibody combination or VAR2 were compared for effectiveness of CTC retrieval. For one patient, enough blood was available to perform the VAR2 beads-based isolation in duplicate. The chart shows the count of isolated CTCs and the fold increase in VAR2 bead-based isolation compared to EpCAM bead-based isolation. (D) Comparison of triple-antibody cocktail-coated beads and VAR2-coated beads for isolating CTCs from patients with PDAC. The left graph illustrates CTC retrieval numbers, the middle graph presents the count of contaminating white blood cells (WBCs), and the right panel displays representative immunostainings ( n = 20). (E) VAR2-based retrieval of CD45 – CK + CTCs for individuals with various conditions, including healthy controls, pancreatitis, intraductal papillary mucinous neoplasms (IPMNs), different stages of PDAC (resectable local, locally advanced, and advanced), neuroendocrine tumors (NETs), and bile duct tumors (cholangiocarcinoma). Boxplots display median values as horizontal lines, with whiskers indicating the 95% CI. (F) Kaplan Meier analysis for overall survival in patients with PDAC patients ( n = 71), dichotomized by the “median CTC numbers” (65 CTCs). Statistical analysis by log rank test. (G) Assessment of the sensitivity of the VAR2-based retrieval method versus serum CA 19-9 levels for detecting local PDAC (left) and non-metastatic PDAC (right), respectively. The upper limit of normal (ULN) of 37 U/mL for CA 19-9 is highlighted in green. In one patient, no CTCs were detected (highlighted in orange). (H) Cox proportional hazards survival regression (Cox regression) analysis for overall 1-year survival. Hazard ratios and 95% confidence interval (CI) are shown. Definition of variables: CTC numbers ≥65/5 mL, disease stage I–IV, gender (M/F), age >65 years, CA 19.9 > 37 U/mL, C-reactive protein >3 mg/L. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001. Please also see Figure S1 .

Article Snippet: PDAC cells (1×10 5 in 50 μL Matrigel) were implanted subcutaneously into 6-8-week-old BALB/c nude mice (Charles River) as described previously., , Once PDAC tumors had reached ∼150 mm 3 , mice were randomly assigned to treatments as outlined in E. Mice were housed according to the institutional guidelines, and the procedures were approved by local animal ethics committees, i.e., SJTU approval n° A-2020-004 and Italian Ministry of Health approval n° 942/2021-PR.

Techniques: Comparison, Derivative Assay, Expressing, Isolation, Cell Culture, Cell Isolation

Journal: Cell Reports Medicine

Article Title: CTC-derived pancreatic cancer models serve as research tools and are suitable for precision medicine approaches

doi: 10.1016/j.xcrm.2024.101692

Figure Lengend Snippet: CTC-derived cultures are more aggressive compared to PDX-derived cultures (A) Invasion assay with fetal bovine serum as chemoattractant for comparing CTC-derived PDAC models versus PDX-derived cultures. Photographs on the left show invaded cells through a Matrigel-coated Boyden chamber, stained with crystal violet. Quantification is presented on the right. Boxplots show the median as a line within the box and use whiskers to represent the 95% CI. (B) In vivo metastasis assay involved injecting 100,000 luciferase-positive (Luc + ) CTC- versus PDX-derived cells into the spleen, followed by splenectomy on day 7 and subsequent tracking of Luc + liver metastasis until day 28. Quantification displayed in the upper-left panel. CTC numbers per mouse were assessed using the VAR2-coated beads method (upper-right panel). Ex vivo liver imaging, including quantification and representative ex vivo liver samples, is shown in the lower panel. (C) Immunohistochemical analysis for cytokeratin 19 (brown) to identify metastases in the liver and lungs of mice, injected with CTC-derived and PDX-derived PDAC cells. (D) Response of six representative CTC-derived models to gemcitabine (GEM), nab-paclitaxel (Abraxane; ABX), or their combination, as assessed by in vitro 3D sphere formation. (E) Corresponding patient’s clinical response for their individually allocated treatment regimen. ∗ p < 0.05 and ∗∗∗ p < 0.001. Please also see Figure S3 .

Article Snippet: PDAC cells (1×10 5 in 50 μL Matrigel) were implanted subcutaneously into 6-8-week-old BALB/c nude mice (Charles River) as described previously., , Once PDAC tumors had reached ∼150 mm 3 , mice were randomly assigned to treatments as outlined in E. Mice were housed according to the institutional guidelines, and the procedures were approved by local animal ethics committees, i.e., SJTU approval n° A-2020-004 and Italian Ministry of Health approval n° 942/2021-PR.

Techniques: Derivative Assay, Invasion Assay, Staining, In Vivo, Luciferase, Ex Vivo, Imaging, Immunohistochemical staining, Injection, In Vitro

Journal: Cell Reports Medicine

Article Title: CTC-derived pancreatic cancer models serve as research tools and are suitable for precision medicine approaches

doi: 10.1016/j.xcrm.2024.101692

Figure Lengend Snippet: CTC-derived cultures are functionally heterogeneous (A) Gating for FACSorting of SiC002 CTC-derived PDAC cells based on their mitochondrial membrane potential (ΔΨm) using tetramethylrhodamine methyl ester (TMRM; left panel). The right panel shows the oxygen consumption rate (OCR) indicative of mitochondrial respiration in ΔΨm high cells versus low cells after sorting. (B) Mitochondrial membrane potential (ΔΨm) stained with TMRM in CSC-enriched sphere versus adherent cultures derived from a diverse set of CTC- and PDX-derived models, respectively ( n = 5). (C) ΔΨm stained with TMRM in CD133 + CSCs versus CD133 – differentiated cancer cells; n = 3. (D) qPCR analysis for mRNA levels of stemness-associated genes in ΔΨm high versus low cells sorted from SiC002 and SiC007 CTC-derived models; n = 4 (unpaired two-tailed t test). (E) Representative gating strategy of PDAC cells with low, medium, and high ΔΨm levels for FACSorting. (F) Representative images (left) and quantification (right) of sphere formation capacity on day 7 following ΔΨm sorting in two different CTC-derived PDAC models; n = 6 (unpaired two-tailed t test). (G) In vivo tumorigenicity of graded cell numbers for two different CTC-derived PDAC models after sorting for ΔΨm levels. Images of explanted tumors (left) and extreme limiting dilution analysis (right). (H) ΔΨm stained with TMRM for CXCR4 + versus CXCR4 – cells. Representative flow cytometry plots of CXCR4 gating and TMRM staining (left), quantification of mean fluorescence intensity (MFI) of TMRM staining (right); n = 3. (I) Immunofluorescence for CXCR4 (white) and TMRM (red) staining. DAPI serves as a nuclear stain. Representative images are on the left, with quantification on the right for two different CTC-derived PDAC models; n = 41 to 208 cells (horizontal lines indicate the mean values). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Please also see Figure S4 .

Article Snippet: PDAC cells (1×10 5 in 50 μL Matrigel) were implanted subcutaneously into 6-8-week-old BALB/c nude mice (Charles River) as described previously., , Once PDAC tumors had reached ∼150 mm 3 , mice were randomly assigned to treatments as outlined in E. Mice were housed according to the institutional guidelines, and the procedures were approved by local animal ethics committees, i.e., SJTU approval n° A-2020-004 and Italian Ministry of Health approval n° 942/2021-PR.

Techniques: Derivative Assay, Membrane, Staining, Two Tailed Test, In Vivo, Flow Cytometry, Fluorescence, Immunofluorescence

Journal: Cell Reports Medicine

Article Title: CTC-derived pancreatic cancer models serve as research tools and are suitable for precision medicine approaches

doi: 10.1016/j.xcrm.2024.101692

Figure Lengend Snippet: Pancreatic CTCs comprise a subset of CXCR4 + CSCs (A) VAR2-based retrieval of CXCR4 + CD45 – CK + CTCs from individuals with various conditions, including healthy controls, pancreatitis, IPMNs, different stages of PDAC (resectable local, locally advanced, and advanced), NETs, and bile duct tumors (cholangiocarcinoma). The content of CD45 – CK + CXCR4 + CTCs among total CTCs at baseline is shown (left). Boxplots indicate median values with horizontal lines, and whiskers represent the 95% CI. Representative image of a retrieved CD45 – CK + CXCR4 + CTC (right). (B) Kaplan-Meier analysis for overall survival in patients with PDAC dichotomized for the “median of CXCR4 + CTC content” (8.9%). Statistical analysis by log rank test. (C) Cox proportional hazards survival regression (Cox regression) analysis for overall 1-year survival, including the variable “CXCR4 + CTC content.” Hazard ratios and 95% CI are shown on a logarithmic scale. (D) Single-cell droplet digital PCR (ddPCR) analysis. Upper panel: schematic illustration of the workflow. Lower panel: ddPCR results of freshly retrieved CXCR4 + versus CXCR4 – CTCs for the indicated genes plotted ( n = 9 each). (E) Flow cytometry illustrating the content of CXCR4 + cells for PDX- versus CTC-derived cultures. (F) Invasive capacity of CXCR4 + versus CXCR4 – cells for CTC- and PDX-derived cultures, respectively. Cells sorted for CXCR4 were placed in the upper chamber of a Boyden chamber, and invasion toward 20% fetal bovine serum (FBS) in the lower chamber was recorded ( n = 4–8). (G) In vivo tumorigenicity of CXCR4 + versus CXCR4 – cells for two representative CTC-derived models. Graded doses (1 to 10 3 cells/injection) were transplanted into immunocompromised nude mice. Table providing quantification (upper panel), estimate of stem cell frequency in vivo for CXCR4 – (black) versus CXCR4 + (red) CTC-derived PDAC cells by limiting dilution assay (lower panel). Depicted is a log-fraction plot of the limiting dilution model. The slope of the line is the log-active cell fraction. The dotted lines give the 95% CI. Analysis was performed at https://bioinf.wehi.edu.au/cgi-bin/limdil/limdil.pl . ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Please also see Figure S5 .

Article Snippet: PDAC cells (1×10 5 in 50 μL Matrigel) were implanted subcutaneously into 6-8-week-old BALB/c nude mice (Charles River) as described previously., , Once PDAC tumors had reached ∼150 mm 3 , mice were randomly assigned to treatments as outlined in E. Mice were housed according to the institutional guidelines, and the procedures were approved by local animal ethics committees, i.e., SJTU approval n° A-2020-004 and Italian Ministry of Health approval n° 942/2021-PR.

Techniques: Digital PCR, Flow Cytometry, Derivative Assay, In Vivo, Injection, Limiting Dilution Assay

Journal: eLife

Article Title: Quantification of microenvironmental metabolites in murine cancers reveals determinants of tumor nutrient availability

doi: 10.7554/eLife.44235

Figure Lengend Snippet: ( A ) Diagram of experimental models used to test the effect of tumor tissue of origin on TIF metabolite levels. Principal component analysis ( B ) and hierarchical clustering ( C ) of PDAC subcutaneous allograft TIF and LUAD subcutaneous allograft TIF samples based on LC/MS measurements of 104 metabolite concentrations. ( D ) Volcano plot depicting the log 2 fold change in metabolite concentration between PDAC and LUAD TIF for metabolites measured using stable isotope dilution. A fold change of 1.5 and raw p-value of 0.01 assuming unequal variance were used to select significantly altered metabolites indicated in pink. For all panels, n = 5 for PDAC subcutaneous allograft TIF samples and n = 10 for LUAD subcutaneous allograft TIF samples. 10.7554/eLife.44235.034 Figure 5—source data 1. Concentrations of 104 metabolites determined by both external standard calibration and stable isotope dilution in all subcutaneous PDAC and LUAD TIF samples in .

Article Snippet: For subcutaneous xenograft studies, 12 week old C57BL/6J animals purchased from Jackson Laboratories (IMSR Cat# JAX:000664, RRID: IMSR_JAX:000664 ) were injected with 100,000 murine PDAC or LUAD cancer cells (suspended in a volume of 100 μL of Matrigel (Corning, 354234) brought to 10 mg/ml with RPMI-1640 (Corning, 50–020-PC) into the subcutaneous space on the flank of the mice.

Techniques: Liquid Chromatography with Mass Spectroscopy, Concentration Assay, Isotope Dilution

Journal: eLife

Article Title: Quantification of microenvironmental metabolites in murine cancers reveals determinants of tumor nutrient availability

doi: 10.7554/eLife.44235

Figure Lengend Snippet: ( A ) Diagram of experimental models used to test the effect of tumor tissue of origin on TIF metabolite levels. Principal component analysis ( B ) and hierarchical clustering ( C ) of PDAC subcutaneous allograft TIF and LUAD subcutaneous allograft TIF samples based on LC/MS measurements of 67 metabolites quantified using isotope-labeled internal standards. For all panels, n = 5 for PDAC subcutaneous allograft TIF samples and n = 10 for LUAD subcutaneous allograft TIF samples. 10.7554/eLife.44235.035 Figure 5—figure supplement 1—source data 1. Concentrations of 66 metabolites determined by only stable isotope dilution in all subcutaneous PDAC and LUAD TIF samples in .

Article Snippet: For subcutaneous xenograft studies, 12 week old C57BL/6J animals purchased from Jackson Laboratories (IMSR Cat# JAX:000664, RRID: IMSR_JAX:000664 ) were injected with 100,000 murine PDAC or LUAD cancer cells (suspended in a volume of 100 μL of Matrigel (Corning, 354234) brought to 10 mg/ml with RPMI-1640 (Corning, 50–020-PC) into the subcutaneous space on the flank of the mice.

Techniques: Liquid Chromatography with Mass Spectroscopy, Labeling, Isotope Dilution