Journal: Journal of Translational Medicine

Article Title: Hypoxic migrasomes drive colorectal cancer liver metastasis by mediating CD5L + macrophage efferocytosis via NRP2/PROX1 axis

doi: 10.1186/s12967-025-07485-0

Figure Lengend Snippet: Phagocytic activity and immunofluorescence validation of CD5L⁺ macrophages in CRC liver metastases following migrasome treatment. ( A ) UMAP blot showing the expression of migrasome marker TSPAN4 in myeloid subsets. ( B ) Boxplot showing efferocytosis scores across the 10 identified myeloid cell subtypes. ( C ) Violin plots depicting the expression of efferocytosis markers CD300B, MERTK, and CD300D across 10 distinct myeloid cell subtypes. ( D ) UMAP plots displaying the expression patterns of three efferocytosis-associated marker genes specifically enriched in CD5L⁺ macrophages. ( E ) Immunofluorescence staining of tumor tissues from MC38-tumor bearing mice showing colocalization of CD163, CD5L, and the migrasome marker MERTK in both treatment groups. Increased MERTK expression is observed in the hypoxic group, indicating enhanced migrasome targeting of CD5L⁺ macrophages

Article Snippet: Samples were incubated with primary antibodies, CD5L (1:500, 17224-1-AP, Proteintech), CD163 (1:200, ab182422, Abcam), NRP2 (1:250, #3366, CST), PROX1(1:250, sc-81983, Santa cruz), MERTK (200 μg/mL, sc-365499, Santa Cruz).

Techniques: Activity Assay, Immunofluorescence, Biomarker Discovery, Expressing, Marker, Staining

Journal: Journal of Translational Medicine

Article Title: Hypoxic migrasomes drive colorectal cancer liver metastasis by mediating CD5L + macrophage efferocytosis via NRP2/PROX1 axis

doi: 10.1186/s12967-025-07485-0

Figure Lengend Snippet: Migrasomal NRP2 is required for CRC-induced CD5L⁺ macrophage differentiation and efferocytosis. ( A ) RT-qPCR analysis confirming efficient knockdown of NRP2 in MC38 cells under hypoxia. ( B ) Western blot analysis confirming efficient knockdown of NRP2 in MC38 cells under hypoxia. ( C ) Flow cytometry analysis of CD5L⁺ macrophage proportion after treatment with control or NRP2-deficient hypoxic migrasomes from MC38 cells. ( D ) Immunofluorescence assay of efferocytosis by CD5L⁺ macrophages following treatment with control or NRP2-deficient hypoxic migrasomes. F4/80 (green) labels macrophages; PI (red) labels apoptotic tumor cells. Scare bar: 50 μm. ( E ) Quantification of mRNA expression of efferocytosis receptors (MERTK, TYRO3, OLR1, CD36, AXL, and TIM3) in macrophages treated with control or NRP2-deficient migrasomes by RT-qPCR. ( F ) Quantification of protein expression of efferocytosis receptors (MERTK, TYRO3, OLR1, CD36, AXL, and TIM3) in macrophages treated with control or NRP2-deficient migrasomes by Western blot. * p < 0.05, ** p < 0.01

Article Snippet: Samples were incubated with primary antibodies, CD5L (1:500, 17224-1-AP, Proteintech), CD163 (1:200, ab182422, Abcam), NRP2 (1:250, #3366, CST), PROX1(1:250, sc-81983, Santa cruz), MERTK (200 μg/mL, sc-365499, Santa Cruz).

Techniques: Quantitative RT-PCR, Knockdown, Western Blot, Flow Cytometry, Control, Immunofluorescence, Expressing

Journal: Journal of Translational Medicine

Article Title: Hypoxic migrasomes drive colorectal cancer liver metastasis by mediating CD5L + macrophage efferocytosis via NRP2/PROX1 axis

doi: 10.1186/s12967-025-07485-0

Figure Lengend Snippet: NRP2–PROX1 interaction promotes CD5L⁺ macrophage differentiation and enhances efferocytosis. ( A ) Co-immunoprecipitation (Co-IP) assays showing that NRP2 interacts with PROX1 in macrophages under normoxic and hypoxic migrasome-treated conditions. ( B ) Immunofluorescence co-localization images confirming the spatial association between NRP2 (green) and PROX1 (red) in macrophages. Nuclei were counterstained with DAPI (blue). Scale bar, 50 μm. ( C ) Flow cytometry analysis showing the proportion of CD5L⁺ macrophages following NRP2 overexpression and/or PROX1 knockdown. ( D ) RT-qPCR analysis of efferocytosis-related genes (AXL, MERTK, and TYRO3) in macrophages with indicated treatments. ( E ) Immunofluorescence staining of F4/80⁺ macrophages (green) engulfing PI-labeled apoptotic MC38 debris (red). Knockdown of PROX1 suppressed efferocytic activity and attenuated the NRP2-induced enhancement. Scale bar, 50 μm. ( F ) Representative fluorescence images and quantification of transwell assay. Fluorescently labeled CRC cells were co-cultured with macrophages overexpressing NRP2, MERTK-knockdown macrophages, or macrophages with combined NRP2 overexpression and MERTK knockdown, and CRC cell transmigration was assessed using a transwell assay. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Samples were incubated with primary antibodies, CD5L (1:500, 17224-1-AP, Proteintech), CD163 (1:200, ab182422, Abcam), NRP2 (1:250, #3366, CST), PROX1(1:250, sc-81983, Santa cruz), MERTK (200 μg/mL, sc-365499, Santa Cruz).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Immunofluorescence, Flow Cytometry, Over Expression, Knockdown, Quantitative RT-PCR, Staining, Labeling, Activity Assay, Fluorescence, Transwell Assay, Cell Culture, Transmigration Assay

Journal: PLoS Pathogens

Article Title: HER2-mediated enhancement of Ebola virus entry

doi: 10.1371/journal.ppat.1008900

Figure Lengend Snippet: Interaction between HER2 and TYRO3 (A), AXL (B), or MERTK (C) in HEK-293T cells transfected with the indicated combination of expression vectors. Cell lysates were immunoprecipitated with anti-FLAG antibody and then immunoblotted. Data are representative of three independent experiments. IP, immunoprecipitation. WCE, whole-cell extract.

Article Snippet: The open reading frame of TYRO3 (pDONR223-TYRO3; Cat# 23916) and MERTK (pDONR223-MERTK; Cat# 23900) genes were obtained from Addgene.

Techniques: Transfection, Expressing, Immunoprecipitation

Journal: PLoS Pathogens

Article Title: HER2-mediated enhancement of Ebola virus entry

doi: 10.1371/journal.ppat.1008900

Figure Lengend Snippet: (A) Phosphorylation level of HER2 and AKT1 in Huh7.0 cells after knockdown of either TYRO3, MERTK, or both. Cells were transfected with the indicated combination of siRNAs and then on day 3 post-transfection, were infected with EBOVΔVP30 at an MOI of 3.0 for 30 min. The indicated protein expression levels were analyzed by immunoblotting. (B) Phosphorylation level of TYRO3 in the Huh7.0 stable cell line expressing TYRO3 after HER2 knockdown. Cells were transfected with siRNAs targeting HER2 or control siRNA and then on day 3 post-transfection, were infected with EBOVΔVP30 at an MOI of 3.0 for 30 min. Cell lysates were immunoprecipitated with anti-TYRO3 antibody and then immunoblotted. Data are representative of three independent experiments. IP, immunoprecipitation. WCE, whole-cell extract. The graph represents the ratio of tyrosine-phosphorylated TYRO3 to total TYRO3 (pTyr/TYRO3) from three independent experiments. (*) indicates a statistically significant difference ( p value ≤ 0.05) from the control.

Article Snippet: The open reading frame of TYRO3 (pDONR223-TYRO3; Cat# 23916) and MERTK (pDONR223-MERTK; Cat# 23900) genes were obtained from Addgene.

Techniques: Phospho-proteomics, Knockdown, Transfection, Infection, Expressing, Western Blot, Stable Transfection, Control, Immunoprecipitation

Journal: PLoS Pathogens

Article Title: HER2-mediated enhancement of Ebola virus entry

doi: 10.1371/journal.ppat.1008900

Figure Lengend Snippet: HER2 interacts with all members of the TAM (TYRO3, AXL, and MERTK) receptor family to form heterodimers. In TAM receptor-mediated cellular entry, EBOV particles attach to the cell surface by binding to a TAM receptor (e.g., TYRO3) via the serum protein GAS6 (1). HER2 also interacts with the virus-bound TYRO3, forming a HER2/TYRO3/EBOV complex on the cell surface (2). The formation of this complex leads to HER2 phosphorylation (either autophosphorylation or phosphorylation by an as yet unknown cellular factor) and the phosphorylation of TYRO3 by HER2 (3), leading to downstream activation of PI3K/AKT signaling pathway (HER2-mediated AKT1 activation) (4) and subsequent induction of macropinocytosis (5). After internalization of the HER2/TYRO3/EBOV complexes into early endosomes via macropinocytosis (6), the complexes are disassociated, and only EBOV particles are delivered to late the endosomes/lysosomes, where later events such as membrane fusion occur. HER2 and TYRO3 are most likely recycled back to the cell surface.

Article Snippet: The open reading frame of TYRO3 (pDONR223-TYRO3; Cat# 23916) and MERTK (pDONR223-MERTK; Cat# 23900) genes were obtained from Addgene.

Techniques: Binding Assay, Virus, Phospho-proteomics, Activation Assay, Membrane

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: Big data analytics and human microarray reveal the key signaling pathways in atherosclerosis. Big data analytics. ( A – C ) Big data analytics for atherosclerosis with 98881cross analyses for overall signaling and 234 cross analyses in aortic arch based on up-to-date RNA-seq data from humans, mouse and rat. In IPA of Pathways and Lists, atherosclerosis was set as the keywords. Microarray in human atherosclerosis. ( D – E ) The top 50 downregulated or upregulated upstream regulators based on activation of z-score. ( F ) Graphical summary of human microarray data (orange: upregulated; blue: downregulated). QIAGEN Ingenuity Pathway Analysis (IPA: 1-atherosclerosis [carotid atherosclerotic plaque] NA CMP_2gGgljQ5SpJAn) and QIAGEN OmicSoft Land Explorer (OLE) were used to analyze microarray data in carotid atherosclerotic plaque from human patients. RNA-seq big data analytics in human . ( G ) MerTK expression in human diseases specifically in related aortic tissues (n = 427 in total), including normal control, embryo, cardiovascular disease (CVD) and respiratory tract disease (RTD)-related cardiovascular disease. MerTK expression was based on RNA-seq or scRNA-seq and was quantified by Log2 (FPKM + 0.1). Original data of RNA-seq or scRNA-seq for MerTK expression were downloaded from QIAGEN OmicSoft Land Explorer. BioGPS . ( H ) MerTK mRNA expression in human cells derived from BioGPS ( http://biogps.org ). The data were analyzed with GraphPad Prism 9.4.1 and shown as the mean ± SD.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Microarray, Protein-Protein interactions, RNA Sequencing, Activation Assay, Expressing, Control, Derivative Assay

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: Endothelial MerTK deficiency promotes the development of atherosclerosis. ( A ) Body weight measurement in MerTK flox/flox mice and MerTK flox/flox Tie2 Cre mice of atherosclerosis model. ( B – C ) The visible formation of atherosclerotic plaque and H&E staining in aortic arch. After the mice were euthanized, the whole aortas were carefully dissected from surrounding tissue and fixed with 10 % neutral buffered formalin solution. The visible formation of atherosclerotic plaque was acquired by an AmScope Trinocular Stereo Microscope with a 12 MP digital camera. ( D ) Immunostaining for Cav-1 expression in aortic arch from MerTK flox/flox mice and MerTK flox/flox Tie2 Cre mice of atherosclerosis model. Mice were injected with a single dose of AAV8-PCSK9 particles along with a high fat diet for two months.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Staining, Microscopy, Immunostaining, Expressing, Injection

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: Endothelial MerTK deficiency promotes proinflammation response and activates p22 phox while inhibits ApoE expression. ( A – D ) Immunostaining for the expression of IL-1β, MCP-1, ApoE, p22 phox and NF-κB i n aortic arch from MerTK flox/flox mice and MerTK flox/flox Tie2 Cre mice that were injected with a single dose of AAV8-PCSK9 particles along with a high fat diet for two months.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Expressing, Immunostaining, Injection

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: Endothelial MerTK deficiency activates pro-atherosclerotic factors. ( A – D ) Immunostaining for the expression of inflammation markers (TNF-α and IFN-γ), NADPH oxidase subunits (p47 phox and gp91 phox ), and MAPK family (ERK, p38 and JNK) in aortic arch from MerTK flox/flox mice and MerTK flox/flox Tie2 Cre mice that were injected with a single dose of AAV8-PCSK9 particles along with a high fat diet for two months.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Immunostaining, Expressing, Injection

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: Proteomics shows the key signaling pathways in endothelial MerTK-mediated atherosclerosis . ( A ) Protein abundance in comparison of MerTK flox/flox group with MerTK flox/flox Tie2 Cre group. ( B ) Volcano plot illustrating differentially expressed proteins in the aortic arch of MerTK flox/flox Tie2 Cre vs. MerTK flox/flox . Relative protein abundance (log2) plotted against significance level (-log10 P-value), showing downregulated (blue), upregulated (red) or non-differentially expressed proteins (grey). ( C ) Graphical summary of proteomics data (orange: upregulated; blue: downregulated). ( D ) The volcano canonical pathways based on activation of z-score (lower panel). Blue: negative value. Orange: positive value. Grey: no activity pattern. Size is based on the number of genes that overlap the pathway. Big data analytics for 67,629 cross analyses for mitochondrial dysfunction based on IPA data base (upper panel). ( E – F ) The top 50 downregulated or upregulated upstream regulators based on activation of z-score.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Protein-Protein interactions, Quantitative Proteomics, Comparison, Activation Assay, Activity Assay

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: MerTK flox/flox Tie2 Cre group demonstrates aggravated mitochondrial dysfunction compared to MerTK flox/flox group. ( A - B ) Machine learning (ML) disease pathways showing mitochondrial dysfunctions and the protein changes of mitochondrial DNA-related disorder (orange: upregulated; blue: downregulated). ( C – F ) Graphical summary for mitochondrial DNA-related disorder, mitochondrial disorder, mitochondrial cytopathy, and mitochondrial myopathy. Red: increased measurement. Green: decreased measurement. Orange: predicted activation. Bule: predicted inhibition. Glos indicates activity when the opposite of measurement. The lines indicate the predicated relationship (orange: leads to activation; blue: leads to inhibition; yellow: findings inconsistent with state of downstream molecule; grey: effect not predicted). ( G – H ) Activated or inhibited molecules associated with mitochondrial dysfunction.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Activation Assay, Inhibition, Activity Assay

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: Proteomics for causal network analysis shows top 50 changed regulators and microRNAs in aortic arch with MerTK flox/flox Tie2 Cre vs. MerTK flox/flox . ( A ) The top 50 activated upstream regulators and IPA prediction of MAPK family and TGFβ family networks based on activation of z-score. ( B ) The top 50 inhibited upstream regulators and IPA prediction of MAPK inhibitor and SB203580 networks based on activation of z-score. ( C ) The upregulated or downregulated microRNAs in MerTK flox/flox Tie2 Cre group compared to MerTK flox/flox group. Representative IPA prediction micorRNA-218 focusing on MAPK signaling pathway. Upregulated and downregulated proteins are highlighted in red and green, respectively, and the color depth is correlated to the fold change. Orange and blue dashed lines with arrows indicate indirect activation and inhibition, respectively. Yellow and grey dashed lines with arrows depict inconsistent effects and no prediction, respectively.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Activation Assay, Inhibition

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: Multi-comparisons for common signaling between proteomics of MerTK flox/flox Tie2 Cre vs. MerTK flox/flox and shared atherosclerosis projects. ( A ) Upstream regulators in MerTK flox/flox Tie2 Cre vs. MerTK flox/flox compared with other atherosclerosis projects based on activation of z-score. ( B ) The volcano canonical pathways based on activation of z-score. ( C ) The toxicity functions analyses based on activation of z-score. ( D ) Diseases and biological functions analysis based on activation of z-score.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Activation Assay

Journal: Redox Biology

Article Title: Endothelial MerTK impairment accelerates the development of atherosclerosis

doi: 10.1016/j.redox.2025.103861

Figure Lengend Snippet: The axis of miR-218-5p/EC MerTK /MAPK represents a novel mechanism of endothelial MerTK-mediated atherosclerosis. ( A ) Expression of p-JNK, p-p38 and p-ERK in HAECs ( B ) Immunostaining for p-ERK expression in HAECs. ( A - B ) HAECs were transfected with transfected with MerTK CRISPR/Cas9 KO Plasmid or control plasmid at 1 μg/6-well in 1 mL for 48 h. Then HAECs were incubated with apoptotic Jurkat cells at a 1:1 apoptotic cell/EC ratio. ( C ) Expression of p-JNK, p-p38 and p-ERK in HAECs. Cells were pre-transfected with miR-218–5p or miR control at 100 nM for 48 h, treated with MerTK inhibitors of UNC 569 or UNC 5293 at 500 nM for 4 h, and then were incubated with apoptotic cells at a 1:1 apoptotic cell/EC ratio for 1 h.

Article Snippet: The MerTK flox/flox mice and Tie2-Cre mice on the C57BL/6J background were generated by Cyagen US (Santa Clara, CA) and housed in the Division of Laboratory Animal Medicine at our institution with an approved ethic number of A23050.

Techniques: Expressing, Immunostaining, Transfection, CRISPR, Plasmid Preparation, Control, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Biosynthesis, Characterization, and Efficacy in Retinal Degenerative Diseases of Lens Epithelium-derived Growth Factor Fragment (LEDGF 1–326 ), a Novel Therapeutic Protein *

doi: 10.1074/jbc.M112.441618

Figure Lengend Snippet: LEDGF1–326 increases phagocytic activity of retinal cells. ARPE-19 cells were transiently transfected with 20 pm/ml MERTK siRNA. Uptake of 2-μm FluoSpheres (100 μg/ml) was monitored to measure the phagocytic activity. LEDGF1–326 increased the uptake of FluoSpheres significantly in a dose-dependent manner, indicating an increase in phagocytic activity of ARPE-19 cells. Data are represented as mean ± S.D. for n = 4. p < 0.05 as compared with the MERTK siRNA-transfected group.

Article Snippet: Phagocytic Assay ARPE-19 cells were seeded in 24-well plates and transfected with 20 p m /ml MERTK siRNA (Santa Cruz Biotechnology Inc., Dallas, TX), using siRNA transfecting agent (Santa Cruz Biotechnology) for 6 h. The transfecting medium was removed, and cells were further incubated in serum-free medium for 24 h. Cells transfected only with the transfecting medium and no MERTK siRNA were maintained as control.

Techniques: Activity Assay, Transfection