polyclonal anti axl antibody  (R&D Systems)

Journal: Metabolism Open

Article Title: Non-coding RNAs in neurodegeneration: an axis-based, evidence-tiered mechanistic synthesis

doi: 10.1016/j.metop.2026.100458

Figure Lengend Snippet: The primary roles of circRNAs. ( A ) circRNAs may function as sponges for miRNAs, as shown by circ-AXL, which has target sites for miR-328. ( B ) circRNAs may engage with certain mRNAs and modulate their stability and/or translation. An instance is the circ-Bptf interaction with p62 mRNA. ( C ) CircRNAs may be translated to yield tiny peptides, as shown by the translation of circCDYL2 into the Cdyl2-60aa peptide, which is around 7 kDa. ( D ) CircRNAs with motifs that can bind to RNA-binding proteins have the ability to function as decoys or sponges for these proteins, therefore regulating their activity. CircRNAs with patterns that promote the interaction between an enzyme and its substrate may function as scaffolds, facilitating the co-localization of the two entities and enhancing reaction kinetics. CircRNAs may engage with gene promoters, recruit TET1 demethylase, and induce substantial demethylation of CpG islands in the DNA. CircRNAs may bind to U1 snRNP and then interact with the RNA polymerase II transcription complex, hence increasing protein expression. snRNA refers to small nuclear RNA, whereas snRNP denotes small nuclear ribonucleoprotein.

Article Snippet: Proteostasis/Protein aggregation , AD , circRNA , circ-AXL, circ-LPAR1 , Modulate inflammatory & amyloid pathways via miRNA axes , Mouse/Cell , Expression + perturbation , Tier 2 , Therapeutic target preclinical , [ , ] .

Techniques: RNA Binding Assay, Activity Assay, Expressing

Journal: Cancer Medicine

Article Title: TYRO3 , AXL , MERTK and Their Ligands in Brain Metastases From Colorectal Cancers

doi: 10.1002/cam4.71869

Figure Lengend Snippet: TAM receptors and ligands' protein expression profile in in vitro models of CRC and orthotopic in vivo model of BM‐CRC. (A) RNAseq data from cell lines presented per cancer type in nTPM (normalized number of transcripts per million) from the Human Protein Atlas database. (B) RNAseq data per colorectal cancer cell line in nTPM from the Human Protein Atlas. (C–G) RNAseq data for PROS1 (C), GAS6 (D), TYRO3 (E), AXL (F), and MERTK (G) in CL40, HT29, LS1034, SW1463, BM‐SC‐CRC1, and BM‐SC‐CRC2 cells. (H–N) Nude mouse brains injected with BM‐SC‐CRC1/2 cells, perfused with 4% PFA, and cut with a microtome (40 μm slices). The control condition (H) is in the absence of a primary antibody showing a unique lesion formed after injection. The red frames correspond to the fields of the higher magnifications (scale bar: Whole brain 2 mm, cropped image 50 μm). Immunostaining for TYRO3 (I), MERTK (J), PROS1 (K) and GAS6 (L) receptors and their ligands (×20 objective, Olympus VS120 slide scanner). The frontier between the human tumor cells and the mouse brain tissue is drawn by a dotted line (*: mouse brain; scale bar: 50 μm). Immunostaining for AXL without (M) and without (N) primary antibody. Scale bar: 50 μm. Nuclei were counterstained in blue.

Article Snippet: The primary antibodies used in our assays were directed against TYRO3 (rabbit polyclonal; #NBP2‐23725; Novus, Bio‐Techne; dilution 1:2000), AXL (goat polyclonal; #AF154; R&D Systems, Bio‐Techne; 1:400), MERTK (rabbit polyclonal; #MKT‐121AP; FabGennix; 1:800), PROS1 (rabbit polyclonal; #A0384; Dako; 1:800), and GAS6 (rabbit polyclonal; generous gift from Dr. Michael Hall [ ]; 1:2000).

Techniques: Expressing, In Vitro, In Vivo, RNA sequencing, Injection, Control, Immunostaining

Journal: Cancer Medicine

Article Title: TYRO3 , AXL , MERTK and Their Ligands in Brain Metastases From Colorectal Cancers

doi: 10.1002/cam4.71869

Figure Lengend Snippet: TAM receptors are expressed in a cohort of CRC patients with BM. Three micrometer paraffin slices of a tissue micro‐array from a cohort of 87 patients with CRC were immunostained for TYRO3, AXL, and MERTK ( n = 87, including n = 37 with BM; [ , , ]; blue Mayer's hematoxylin counterstaining). (A–C) Immunostaining scores were assigned from 1 to 4, according to the percentage of positive tumor cells, as shown in these representative images of TYRO3 (A), AXL (B), and MERTK (C) staining in biopsies from the tumor center. Arrows highlight blood vessels (scale bar: 300 μm; ×20 magnification on Olympus VS120 slide scanner). (D) Evolution of the immunostaining score from the primary tumor to the BM.

Article Snippet: The primary antibodies used in our assays were directed against TYRO3 (rabbit polyclonal; #NBP2‐23725; Novus, Bio‐Techne; dilution 1:2000), AXL (goat polyclonal; #AF154; R&D Systems, Bio‐Techne; 1:400), MERTK (rabbit polyclonal; #MKT‐121AP; FabGennix; 1:800), PROS1 (rabbit polyclonal; #A0384; Dako; 1:800), and GAS6 (rabbit polyclonal; generous gift from Dr. Michael Hall [ ]; 1:2000).

Techniques: Microarray, Immunostaining, Staining

Journal: Cancer Medicine

Article Title: TYRO3 , AXL , MERTK and Their Ligands in Brain Metastases From Colorectal Cancers

doi: 10.1002/cam4.71869

Figure Lengend Snippet: Prognostic value of TAM receptors in mCRC TCGA and BM‐CRC cohorts. Kaplan–Meier survival curves (A–C). TAM in the TCGA cohort of 85 patients with metastatic disease upon diagnosis. OS is defined as the time from primary tumor diagnosis. Patients were split according to the RNA expression level in the tumor (RSEM) into Low or High. Overall survival depending on the combined expression of TYRO3‐GAS6 (A), AXL‐GAS6 (B), MERTK‐GAS6 (C). (D–F) In the BM‐CRC cohort, patients were split according to the staining score in immunohistochemistry into Low (0, 1, 2) or High (3, 4). OS is the time from metastatic disease diagnosis; TYRO3 in the primary tumor (D), AXL in the primary tumor (E), MERTK in the primary tumor (F). (G–I) TYRO3 in BM (G), AXL in BM (H), MERTK in BM (I).

Article Snippet: The primary antibodies used in our assays were directed against TYRO3 (rabbit polyclonal; #NBP2‐23725; Novus, Bio‐Techne; dilution 1:2000), AXL (goat polyclonal; #AF154; R&D Systems, Bio‐Techne; 1:400), MERTK (rabbit polyclonal; #MKT‐121AP; FabGennix; 1:800), PROS1 (rabbit polyclonal; #A0384; Dako; 1:800), and GAS6 (rabbit polyclonal; generous gift from Dr. Michael Hall [ ]; 1:2000).

Techniques: Biomarker Discovery, RNA Expression, Expressing, Staining, Immunohistochemistry

Journal: Cancer Medicine

Article Title: TYRO3 , AXL , MERTK and Their Ligands in Brain Metastases From Colorectal Cancers

doi: 10.1002/cam4.71869

Figure Lengend Snippet: Graphical abstract. TAM receptors and their ligands in brain metastases from colorectal cancers. We assessed TAM receptors in different models. Created with Biorender.com . First, stem‐cell enriched BM‐derived cell lines from two CRC patients, CRC1 and CRC2. Orthotopic BM mice model and RNAseq data were generated, showing that AXL and its ligand GAS6 were poorly expressed in those stem‐cell enriched models. Second, we stained for TAM receptors on tissues from the primary tumor and metastatic sites in CRC1 and CRC2 patients, discovering that AXL was expressed in those tissues but on endothelial cells. TYRO3 was instead expressed on tumor cells and MERTK on leukocytes. Third, we stained for TAM receptors in a tissue micro‐array of primary tumor, non‐brain, and brain metastases from a local cohort of 85 patients. This confirmed the vascular location of AXL and showed the evolution of TAM protein expression from primary tumor to BM tissue. Last, we studied the prognostic properties of TAM receptors and their ligand in a TCGA cohort of patients with metastatic CRC. GAS6 appeared as a robust prognostic factor and suggested a prognostic impact of the balance between AXL and GAS6 expression as patients with tumors expressing Low AXL/High GAS6 had a significantly shortened overall survival. Finally, we provide a comprehensive study of TAM expression in metastatic CRC with a strong focus on BM‐CRC, underlining AXL expression evolution from primary tumor to BM and its prognostic potential in BM‐CRC. Although non‐significant, AXL proteic expression stratifies better the patients with BM‐CRC when assessed in brain tissue instead of primary tumor tissue. AS compared with TYRO3 and MERTK, AXL proteic expression is more frequently conserved, meaning increased or stable, from primary tumor to brain metastasis.

Article Snippet: The primary antibodies used in our assays were directed against TYRO3 (rabbit polyclonal; #NBP2‐23725; Novus, Bio‐Techne; dilution 1:2000), AXL (goat polyclonal; #AF154; R&D Systems, Bio‐Techne; 1:400), MERTK (rabbit polyclonal; #MKT‐121AP; FabGennix; 1:800), PROS1 (rabbit polyclonal; #A0384; Dako; 1:800), and GAS6 (rabbit polyclonal; generous gift from Dr. Michael Hall [ ]; 1:2000).

Techniques: Derivative Assay, RNA sequencing, Generated, Staining, Microarray, Expressing

Journal: NPJ Precision Oncology

Article Title: AXL–SHC1 signaling axis mediates adaptive resistance to HER2-targeted tyrosine kinase inhibitors in HER2-aberrant lung and gastric cancers

doi: 10.1038/s41698-026-01385-2

Figure Lengend Snippet: Cell viability of A Calu-3 and B MKN7 cells was assessed after incubation with mobocertinib (0.01 or 0.1 μmol/L) in the presence or absence of the AXL inhibitor ONO7475 (0.3 μmol/L) for 72 h using MTT assays. Western blotting analysis of C Calu-3 and D MKN7 cells treated with mobocertinib (0.01 or 0.1 μmol/L) with or without ONO7475 (0.3 μmol/L) for 4 h. E Visualization of Calu-3 and MKN7 cells using crystal violet staining after 14 days of treatment with HER2-TKIs, ONO-7475 (0.3 µmol/L), or a combination of HER2-TKIs and ONO7475. Calu3 cell was treated with 0.01 µmol/L mobocertinib, 0.01 nmol/L poziotinib, 0.1 µmol/L tucatinib, 0.3 μmol/L ONO7475, or a combination of each HER2-TKI and 0.3 μmol/L ONO-7475 for 14 days. MKN7 cells were treated with 0.1 µmol/L mobocertinib, 0.1 nmol/L poziotinib, 1.0 µmol/L tucatinib, 0.3 μmol/L ONO7475, or a combination of each HER2-TKI and 0.3 μmol/L ONO-7475 for 14 days. F Flow cytometry analysis of apoptotic cell percentages in Calu-3 and MKN7 cells treated with mobocertinib (0.01 or 0.1 μmol/L), ONO7475 (0.3 µmol/L), or a combination of mobocertinib and ONO7475 for 48 h. * P < 0.05 (one-way ANOVA).

Article Snippet: The HER2-TKIs mobocertinib, poziotinib, tucatinib, afatinib and sevabertinib, as well as the AXL inhibitors ONO7475 and NPS1034, were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Incubation, Western Blot, Staining, Flow Cytometry

Journal: NPJ Precision Oncology

Article Title: AXL–SHC1 signaling axis mediates adaptive resistance to HER2-targeted tyrosine kinase inhibitors in HER2-aberrant lung and gastric cancers

doi: 10.1038/s41698-026-01385-2

Figure Lengend Snippet: A Calu-3 and MKN7 cells were left untreated (left) or treated with mobocertinib (1.0 μmol/L) for 9 d (drug-tolerant (DT) cells; middle), and DT cells were incubated with a drug-free medium for 9 d (drug-free (DF) cells; right). B Calu-3 and MKN7 parental, DT, and DF cells were treated with the indicated concentration of mobocertinib for 72 h and assessed for cell viability using the MTT assay. C Calu-3 and MKN7 DT cells were incubated with mobocertinib (0.01 or 0.1 μmol/L), ONO7475 (0.3 μmol/L), or a combination of mobocertinib and ONO7475 for 72 h. Cell growth was determined using MTT assays. D Western blotting of Calu-3 and MKN7 DT cells treated with mobocertinib (0.01 or 0.1 μmol/L), ONO7475 (0.3 μmol/L), or a combination of mobocertinib and ONO7475 for 4 h.

Article Snippet: The HER2-TKIs mobocertinib, poziotinib, tucatinib, afatinib and sevabertinib, as well as the AXL inhibitors ONO7475 and NPS1034, were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Incubation, Concentration Assay, MTT Assay, Western Blot

Journal: NPJ Precision Oncology

Article Title: AXL–SHC1 signaling axis mediates adaptive resistance to HER2-targeted tyrosine kinase inhibitors in HER2-aberrant lung and gastric cancers

doi: 10.1038/s41698-026-01385-2

Figure Lengend Snippet: A Calu-3 CDX tumors were treated with vehicle (control), ONO7475 10 mg/kg, mobocertinib 15 mg/kg, or ONO7475 10 mg/kg plus mobocertinib 15 mg/kg ( n = 6, per group). Tumor volumes were measured over time, and the results are shown as the mean ± SEM. Statistical comparisons were conducted using two-way ANOVA. * P < 0.05. B Western blotting analysis of Calu-3 CDX tumors treated with mobocertinib (15 mg/kg), ONO7475 (10 mg/kg), or a combination thereof for 4 days. C Representative immunohistochemistry images of Calu-3 CDX tumors stained with hematoxylin and eosin (HE), specific human Ki-67 and TUNEL. Scale bar, 50 μm. Quantification of proliferating cells determined by D Ki67-positive proliferation index (percentage of Ki67-positive cells) and E TUNEL-positive proliferation index (percentage of TUNEL-positive cells) in Calu-3 CDX tumors. The columns represent the mean of five evaluated areas, and bars represent the standard deviation (SD). Statistical comparisons were conducted using one-way ANOVA. * P < 0.05. F Representative immunohistochemistry images of Calu-3 CDX tumors treated with vehicle (control) or mobocertinib 15 mg/kg for 4 days and stained with specific human ShcBP1 antibodies. Scale bar, 50 μm. G H2170 CDX tumors containing the transfected vector or overexpressing AXL were treated with vehicle (control) or mobocertinib 10 mg/kg ( n = 6, per group) or ONO7475 10 mg/kg plus mobocertinib 10 mg/kg. Tumor volumes were measured over time, and the results are shown as mean ± SEM. Statistical comparisons were conducted using two-way ANOVA. *comparison of therapeutic efficacy of mobocertinib comparing tumors derived from vector-transfected or AXL-overexpressing H2170 cells; **comparison of therapeutic efficacy of the combination of mobocertinib and ONO7475 to mobocertinib monotherapy in AXL-overexpressing H2170 tumors.

Article Snippet: The HER2-TKIs mobocertinib, poziotinib, tucatinib, afatinib and sevabertinib, as well as the AXL inhibitors ONO7475 and NPS1034, were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Control, Western Blot, Immunohistochemistry, Staining, TUNEL Assay, Standard Deviation, Transfection, Plasmid Preparation, Comparison, Drug discovery, Derivative Assay