Journal: medRxiv

Article Title: A DDR2-targeted PET tracer images activated fibroblasts in early pulmonary fibrosis

doi: 10.1101/2025.11.26.25341068

Figure Lengend Snippet: a , Representative longitudinal PET/CT and CT images of unilateral bleomycin (BLM)-induced pulmonary fibrosis in hFAP-knock-in (hFAP-BLM) mice imaged with 18 F-FAPI and in wild-type C57BL/6-BLM mice imaged with 68 Ga-NOTA-1A12 at days 3, 7, 14, 21, and 28 after modeling. White dashed lines delineate the fibrotic lung regions. b , Quantitative analysis of probe uptake in fibrotic lung regions expressed as SUV mean. Data are presented as mean ± SD ( n = 8∼10 per group). Statistical significance was determined using one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns: not significant. c , Schematic diagram of the sequential dual-probe PET/CT imaging protocol in the bleomycin-induced pulmonary fibrosis crab-eating macaque model. The macaque was first imaged with 18 F-FAPI on day 4 post-BLM instillation, followed by 68 Ga-NOTA-1A12 imaging on day 6 after sufficient radioactive decay. d , Representative axial, coronal, and maximum intensity projection (MIP) PET/CT images showing lung fibrotic regions with probe accumulation.

Article Snippet: To generate stable overexpression models, lentiviral vectors carrying human DDR2 or hFAP complementary DNA (IGE Biotechnology, Guangzhou, China) were constructed and co-transfected with packaging plasmids psPAX2 and pMD2.G into 293T cells using Lipofectamine 3000 (Invitrogen).

Techniques: Positron Emission Tomography-Computed Tomography, Knock-In, Imaging

Journal: Cancer Research

Article Title: ANGPTL4 Induction by Prostaglandin E2 under Hypoxic Conditions Promotes Colorectal Cancer Progression

doi: 10.1158/0008-5472.can-11-1262

Figure Lengend Snippet: Figure 1. PGE2 enhances ANGPTL4 expression under hypoxic conditions. A and B, LS- 174T cells were treated with the indicated concentrations of PGE2 and exposed to normoxia/hypoxia for 12 hours (mRNA) or 48 hours (protein) to determine the expression of ANGPTL4. ANGPTL4 mRNA (A) and protein (B) levels were determined by qRT-PCR and ELISA analysis, respectively. C, ELISA analysis was done in HT-29 (left) and HCT-116 (right) cells exposed to normoxia/hypoxia or dimethyl sulfoxide (DMSO)/1 mmol/ L PGE2 for 48 hours. , P < 0.05; , P < 0.01; , P < 0.001.

Article Snippet: ANGPTL4 cDNA (Origene) was amplified with pfuUltra Hotstart DNA polymerase (Stratagene).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Cancer Research

Article Title: ANGPTL4 Induction by Prostaglandin E2 under Hypoxic Conditions Promotes Colorectal Cancer Progression

doi: 10.1158/0008-5472.can-11-1262

Figure Lengend Snippet: Figure 2. EP1 mediates PGE2- induced ANGPTL4 expression during hypoxia. A, LS-174T cells were exposed to hypoxia for 12 hours and qRT-PCR was done to determine the expression of EP1, EP2, EP3, and EP4 receptors (left). LS-174T cells were exposed to hypoxia, and EP1 protein levels were analyzed by Western blotting over times as indicated (right). Actin was used as a protein loading control. B, LS-174T cells were pretreated with EP1 (10 mmol/L SC- 19220) or EP4 (10 mmol/L ONO- AE3-208) antagonists for 1 hour and then were treated with PGE2 under normoxic or hypoxic conditions, following qRT-PCR and ELISA to detect ANGPTL4 mRNA and protein. C, ANGPTL4 levels were determined in the growth medium with a commercial ELISA after treatment with the indicated concentration of EP1 agonist (17-PT-PGE2) under normoxic or hypoxic conditions. D, cells were pretreated with PLC inhibitor (10 mmol/L U73122) or Ca2þ

Article Snippet: ANGPTL4 cDNA (Origene) was amplified with pfuUltra Hotstart DNA polymerase (Stratagene).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Cancer Research

Article Title: ANGPTL4 Induction by Prostaglandin E2 under Hypoxic Conditions Promotes Colorectal Cancer Progression

doi: 10.1158/0008-5472.can-11-1262

Figure Lengend Snippet: Figure 3. ANGPTL4 increases tumor growth in vitro and in vivo. A, LS-174T cells were treated with full- length (F–A) or C-terminal fragment (C–A) of recombinant ANGPTL4 protein for 3 days to determine cell proliferation (top). HCT-116 and HT-29 cells were exposed to 100 ng/mL C-ANGPTL4 for 3 days and cell proliferation was analyzed (bottom). B, LS-174T cells were stably transfected with GFP or ANGPTL4 constructs. C-ANGPTL4 expression was determined by Western blotting after the collection of conditioned medium (top). These cells were incubated in serum-free medium for 3 days and then cell proliferation was tested (bottom). C, 1 mmol/L of PGE2 was treated in LS-174T cells exposed to normoxia/hypoxia for 3 days and cell proliferation assay was done. D, LS-174T cell stably transfected with GFP or ANGPTL4 constructs were injected into the flanks of nude mice. The tumor size (middle) was measured from 13 to 26 days after injection, and tumor weight (right) was also measured and C- ANGPTL4 level (left) was detected after mice were euthanized. , P < 0.05; , P < 0.01; , P < 0.001.

Article Snippet: ANGPTL4 cDNA (Origene) was amplified with pfuUltra Hotstart DNA polymerase (Stratagene).

Techniques: In Vitro, In Vivo, Recombinant, Stable Transfection, Transfection, Construct, Expressing, Western Blot, Incubation, Proliferation Assay, Injection

Journal: Cancer Research

Article Title: ANGPTL4 Induction by Prostaglandin E2 under Hypoxic Conditions Promotes Colorectal Cancer Progression

doi: 10.1158/0008-5472.can-11-1262

Figure Lengend Snippet: Figure 4. ANGPTL4 enhances cancer cell growth through STAT1 A, LS-174T cells were exposed to C-ANGPTL4 for 12 hours (mRNA) and 24 hours (protein) and qRT-PCR and Western blot were done for STAT1 mRNA and protein levels, respectively (top). Cells treated with vehicle (CTL) or C-ANGPTL4 were stained with antibody against STAT1 (green) and counterstained with DAPI (blue). Immunofluorescence was detected by a fluorescence microscope (Nikon ECLIPSE TE300; 400; middle). Immunohistochemistry was done to detect STAT1 expression in tumor tissue generating by injection of ANGPTL4/LS-174 cells into the flanks of nude mice compared with the GFP control (bottom). B, cells were transfected with nontargeting and STAT1 siRNA, respectively. After 24 hours, transfecting cells were treated with C-ANGPTL4 for 3 days and subjected to Western blotting for STAT1 (top) and cell proliferation analysis (bottom). C, Western blot analysis was done in LS-174T cells exposed to normoxia/hypoxia or DMSO/1 mmol/L PGE2 for 24 hours to determine the expression of STAT1 and the phosphorylation of ERK and Src. D, Western blot analysis was carried out to examine the phosphorylation of ERK and Src in LS-174T cells in the presence or absence of C-ANGPTL4 (top). The cells were pretreated with 1 mmol/L SU6656 (SU) or 10 mmol/L PD98059 (PD) for 1 hour and then were treated with C-ANGPTL4. STAT1 levels were determined (middle). Cells were treated with 2 mmol/L of diphenylene iodonium (DP1) and C-ANGPTL4 and then the phosphorylation of ERK and Src (0.5 h) as well as the expression of STAT1 (24 h) were determined (bottom). , P < 0.05; , P < 0.01.

Article Snippet: ANGPTL4 cDNA (Origene) was amplified with pfuUltra Hotstart DNA polymerase (Stratagene).

Techniques: Quantitative RT-PCR, Western Blot, Staining, Microscopy, Immunohistochemistry, Expressing, Injection, Control, Transfection, Phospho-proteomics

Journal: Cancer Research

Article Title: ANGPTL4 Induction by Prostaglandin E2 under Hypoxic Conditions Promotes Colorectal Cancer Progression

doi: 10.1158/0008-5472.can-11-1262

Figure Lengend Snippet: Figure 5. COX-2, ANGPTL4, and STAT1 expressions in colorectal cancers. A, total RNA was isolated from 30 individual human colorectal cancer tissues and corresponding normal mucosa. Equal amounts of mRNA were analyzed by qRT-PCR for COX-2, ANGPTL4, and STAT1. Thecross line indicates a ratio equal to 1. B, colorectal cancer patients in the Moffitt cohort (n ¼ 177) were dichotomized by expression of ANGPTL4 and patient with relative high expression or relative low expression of ANGPTL4 were considered for plotting. DFS, disease-free survival. C, correlation of COX-2 and STAT1 or ANGPTL4 expression in Moffitt colorectal cancer patient cohort. Scatter plots between COX-2 and STAT1 or ANGPTL4 in Moffitt cohort (N ¼ 177).

Article Snippet: ANGPTL4 cDNA (Origene) was amplified with pfuUltra Hotstart DNA polymerase (Stratagene).

Techniques: Isolation, Quantitative RT-PCR, Expressing