vegfr1 flt1  (Proteintech)

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A ) THP-1 cells were infected with SFTSV (MOI = 1) for 12, 24, 36, 48, and 60 h. SFTSV viral RNA (left) and sVEGFR1 mRNA (right) were measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (SFTSV: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001, *** P < 0.0001, *** P < 0.0001; sVEGFR1: *** P = 0.0002, *** P < 0.0001, *** P = 0.0001, *** P < 0.0001, *** P < 0.0001). ( B ) THP-1 cells were infected with SFTSV for 12, 24, 36, 48, and 60 h. Intracellular sVEGFR1 and the viral NP protein were measured by western blotting. sVEGFR1 was stained with a specific monoclonal antibody and GAPDH served as an internal control. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (ns = 0.2035, ** P = 0.0043, *** P = 0.0003, *** P = 0.0002, * P = 0.0127). ( C ) THP-1 cells were infected with SFTSV for 12, 24, 36, 48 and 60 h. The secreted sVEGFR1 in cell supernatant was measured by capture ELISA. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (ND: Not Detected, *** P = 0.0002, ** P = 0.0049, *** P < 0.0001, *** P < 0.0001). ( D – H ) THP-1 cells were infected with SFTSV for 12, 24, 36, 48 and 60 h at various MOIs (MOI = 0.1, 1, and 10). Viral RNA (left) and sVEGFR1 mRNA (right) were determined by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (12 h: left: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001; right: * P = 0.0169, ** P = 0.0087, *** P < 0.0001. 24 h: left: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001; right: *** P = 0.0005, *** P < 0.0001, *** P = 0.0002. 36 h: left: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001; right: *** P = 0.0004, *** P < 0.0001, *** P = 0.0002. 48 h: left: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001; right: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001. 60 h: left: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001; right: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001). ( I – M ) THP-1 cells were infected with SFTSV for 12, 24, 36, 48 and 60 h at various MOIs (MOI = 0.1, 1, and 10). sVEGFR1 in cell supernatant was measured by capture ELISA. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (12 h: ns = 0.0910, ns = 0.4550, *** P < 0.0001; 24 h: ns = 0.8551, *** P = 0.0002, *** P < 0.0001; 36 h: ns = 0.6919, *** P < 0.0001, *** P < 0.0001; 48 h: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001; 60 h: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001). ( N ) Primary human monocyte-derived-macrophages were infected with SFTSV (MOI = 1) for 24 h. SFTSV viral RNA (left) and sVEGFR1 mRNA (right) were measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (SFTSV: *** P < 0.0001; sVEGFR1: *** P = 0.0002). ( O ) Primary human monocyte-derived-macrophages were infected with SFTSV (MOI = 1) for 24 h. The secreted sVEGFR1 in cell supernatant was measured by capture ELISA. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (** P = 0.0026). ( P ) Bone marrow-derived macrophages (BMDMs) from C57BL/6J mice were matured to macrophage and then infected with SFTSV (MOI = 1) for 24 h. BMDMs were infected with SFTSV for 24 h. Viral RNA (left) and sVEGFR1 (right) mRNA were measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (SFTSV: *** P < 0.0001; sVEGFR1: * P = 0.0111). Data information: Data shown are mean ± SD of three biological replicates. (ns, P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001). .

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Infection, Two Tailed Test, Western Blot, Staining, Control, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A ) THP-1 cells were infected with SFTSV for 12, 24, 36, 48, and 60 h. Total VEGFR1 mRNA was measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (ns = 0.0548, *** P < 0.0001, *** P < 0.0001, *** P < 0.0001, *** P < 0.0001) ( B ) THP-1 cells were infected with SFTSV for 12, 24, 36, 48, and 60 h. Then the cells were incubated with anti-VEGFR1 for 1 h at 4 °C and VEGFR1 level on the cell membrane was determined by flow cytometry. X-axes show anti-VEGFR1 antibody (logarithm of fluorescence), Y axes depict the cell count. ( C , D ) THP-1 cells were induced to differentiate into macrophage-like cells by 150 nM phorbol-12-myristate-13-acetate (PMA) and then infected with SFTSV. Total VEGFR1 ( C ) and sVEGFR1 ( D ) mRNAs were measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (( C) : *** P < 0.0001; ( D ): *** P < 0.0001). ( E ) Primary human monocyte-derived-macrophages were infected with SFTSV (MOI = 1) for 24 h. Total VEGFR1 mRNA was measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (* P = 0.0198). Data information: Data shown are mean ± SD of three biological replicates. (ns, P > 0.05; * P < 0.05; *** P < 0.001).

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Infection, Two Tailed Test, Incubation, Membrane, Flow Cytometry, Fluorescence, Cell Counting, Derivative Assay

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A , B ) THP-1 cells were induced to differentiate into M1 macrophage-like cells by 24 h incubation with 150 nM PMA, followed by IFN-γ (20 ng/ml) and LPS (10 pg/ml), or THP-1 cells were induced to differentiate into M2 macrophage-like cells by 24 h incubation with 150 nM PMA, followed by interleukin 4 (20 ng/ml) and interleukin 13 (20 ng/ml) for 72 h. Total VEGFR1 ( A ) and sVEGFR1 ( B ) mRNAs were measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (( A ): *** P < 0.0001; ( B ): *** P < 0.0001). ( C , D ) Primary human monocyte-derived-macrophages were differentiated into M1 macrophages by culture in the presence of IFN-γ (20 ng/ml) and LPS (20 ng/ml) for 24 h, or were differentiated into M2 macrophages by culture in the presence of interleukin 4 (20 ng/ml) and interleukin 13 (20 ng/ml) for 72 h. Total VEGFR1 ( C ) and sVEGFR1 ( D ) were measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (( C ): ** P = 0.0036; ( D ): *** P < 0.0001). ( E ) Primary human monocyte-derived-macrophages were differentiated into M1 macrophages or M2 macrophages. The secreted sVEGFR1 in cell supernatant was measured by capture ELISA. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (** P = 0.0011). ( F ) Primary human monocyte-derived-macrophages were differentiated into M1 macrophages or M2 macrophages. EGR1 mRNA was measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (** P = 0.0018). Data information: Data shown are mean ± SD of three biological replicates. (** P < 0.01; *** P < 0.001).

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Incubation, Two Tailed Test, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A ) hu-PBL NCG mice were infected with SFTSV ( n = 5) at 1 × 10 3 TCID 50 or mock-infected ( n = 5), and blood samples were collected after 14 days. SFTSV viral RNA (left) and sVEGFR1 mRNA (right) in blood cells were determined by qPCR. n = 5 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (SFTSV, *** P < 0.0001; sVEGFR1, *** P = 0.0001). ( B ) The serum level of sVEGFR1 protein was measured by capture ELISA. Each point represents an individual. n = 5 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (*** P = 0.0008). ( C ) The infected mice ( n = 5) were euthanized, and spleens were collected. sVEGFR1 and the viral NP protein in spleens were measured by western blotting (left). Numeric label indicates individual mice. The sVEGFR1 protein expression relative to GAPDH was shown (right). n = 3 biological replicates for Mock; n = 5 biological replicates for SFTSV. Statistical significance was determined by two-tailed unpaired Student t test. (** P = 0.0074). ( D ) Detection of sVEGFR1 by immunofluorescence in the spleens after SFTSV infection (left). SFTSV-Gn (red), sVEGFR1 (green) and nuclei (DAPI, blue) were visualized using either antibodies specific for Gn or sVEGFR1, or DAPI for nuclei. A merged image of red, green and blue channels is shown (merge). The quantifications of relative sVEGFR1 intensities were determined by ImageJ software (right). Scale bars, 25 µm, respectively. n = 5 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (*** P < 0.0001). ( E ) Blood samples were obtained from 60 SFTS patients (grouped into severe and mild based upon clinical criteria) and 30 healthy individuals as controls. The serum level of sVEGFR1 was measured by capture ELISA. n = 30 biological replicates for each group. Statistical significance was determined by two-tailed unpaired Student t test. (Mild, *** P < 0.0001; Severe, *** P < 0.0001). Data information: Data shown are mean ± SD of at least three biological replicates with each data point representing a biological experiment (** P < 0.01; *** P < 0.001). .

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Infection, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Immunofluorescence, Software

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A ) HUVECs were introduced into the Matrigel in the presence of serum. Each serum collected from SFTS patients of either mild symptom or severe symptom was placed under UV light for 30 min for residual virus inactivation. Tube formation of HUVECs was examined at 12 h after treatment. Representative capillary tubule structures were shown (top). Red arrowheads indicate tube elongation. Bar graph (bottom) represent the fold change of tubule formation. Scale bar = 50 μm. n = 3 biological replicates for each group. Statistical significance was determined by two-tailed unpaired Student t test. (Mild, *** P < 0.0001; Severe, *** P < 0.0001). ( B ) Representative images (top) of mouse aortic rings from which microvessels sprouted after treatment with sera collected from either mild patients or severe patients. Each serum was placed under UV light for 30 min for residual virus inactivation. Red arrowheads indicate micro-vascular sprouts. Bar graph (bottom) represent the fold change of microvessels outgrowth from aortic rings. Scale bar = 50 μm. n = 3 biological replicates for each group. Statistical significance was determined by two-tailed unpaired Student t test. (Day 3: * P = 0.0281, * P = 0.0281; Day 5: *** P < 0.0001, *** P < 0.0001). ( C ) Sera from healthy individuals and SFTS patients were placed under UV light for 30 min to inactivate the virus, and then pre-treated with a sVEGFR1 neutralizing antibody-conjugated or control antibody-conjugated magnetic beads to remove sVEGFR1. HUVECs were introduced into the Matrigel in the presence of pre-treated-serum from either healthy individuals or SFTS patients, and tube formation of HUVECs was examined at 12 h after treatment. Representative capillary tubule structures were shown. Red arrowheads indicate tube elongation. Scale bar = 50 μm. Bar graph on the right represent the fold change of tubule formation. n = 3 biological replicates for each group. Statistical significance was determined by two-tailed unpaired Student t test. (Control, ** P = 0.0065; SFTS, * P = 0.0177). ( D ) Representative images of the tube formation following incubation with pre-treated sera from either healthy individuals or SFTS patients added with exogenous recombinant sVEGFR1 (10 ng/ml) or VEGFA (20 ng/ml) protein, respectively. Red arrowheads indicate tube elongation. Scale bar = 50 μm. Bar graph on the right represent the fold change of tubule formation. n = 3 biological replicates for each group. Statistical significance was determined by two-tailed unpaired Student t test. (12 h: ** P = 0.0041, ** P = 0.0021; 24 h: ** P = 0.0030, ** P = 0.0022). Data information: Data shown are mean ± SD of three biological replicates. (* P < 0.05; ** P < 0.01; *** P < 0.001). .

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Virus, Two Tailed Test, Control, Magnetic Beads, Incubation, Recombinant

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A ) HeLa cells were transiently transfected with a plasmid expressing EGR1 or an empty vector (EV-his). After 36 h post transfection, the relative EGR1 mRNA (left) and sVEGFR1 mRNA (right) in HeLa cells were measured by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (EGR1: *** P < 0.0001; sVEGFR1: *** P < 0.0001). ( B ) The expression of sVEGFR1 was measured by western blotting (left) and quantified by density of the WB bands (right). n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (*** P = 0.0008). ( C ) sVEGFR1 protein level in HeLa cell culture supernatant was measured by ELISA. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (** P = 0.0013). ( D ) HEK-293T cells were infected with a recombinant lentivirus vector (pLV-Puro-EGR1) and screened with puromycin for cell clones with stable EGR1 overexpression (EGR1-293T cells). EGR1-293T cells, along with HEK-293T cells infected with pLV-Puro vector (Con-293T cells), were tested for the mRNA levels of EGR1 and sVEGFR1 by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (EGR1: *** P < 0.0001; sVEGFR1: ** P = 0.0050). ( E ) The protein levels of EGR1 and sVEGFR1 in EGR1-293T cells and Con-293T cells were measured by western blotting (left) and quantified by density of the WB bands (right). n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (*** P = 0.0003). ( F ) sVEGFR1 protein in cell culture supernatant was measured by ELISA. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (*** P < 0.0001). ( G ) Cells transfected with siNC or siEGR1 (siEGR1-1, siEGR1-2 and siEGR1-3) were harvested at 36 hpt to evaluate EGR1 protein knock-down efficiency and sVEGFR1 protein levels by western blotting. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (EGR1: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001; sVEGFR1: *** P < 0.0001, *** P < 0.0001, *** P < 0.0001). Data information: Data shown are mean ± SD of three biological replicates. (** P < 0.01; *** P < 0.001). .

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Transfection, Plasmid Preparation, Expressing, Two Tailed Test, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Infection, Recombinant, Clone Assay, Over Expression, Knockdown

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A, B ) THP-1 cells transfected with siNC or siEGR1 (siEGR1-2 in Fig. ) were harvested at 36 hpt to evaluate EGR1 mRNA ( A ) knock-down efficiency and sVEGFR1 mRNA ( B ) levels by qPCR. n = 3 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (( A ): *** P = 0.0003; ( B ): * P = 0.0243). Data information: Data shown are mean ± SD of three biological replicates (* P < 0.05; *** P < 0.001).

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Transfection, Knockdown, Two Tailed Test

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A ) Schematic representation of experimental design. The Matrigel plug assays were performed by subcutaneous injection of Matrigel containing sVEGFR1 Ab or Control Ab into hu-PBL NCG mice followed with or without SFTSV infection. Seven days after infection, the Matrigel plugs were removed and photographed. ( B ) Images of Matrigel plugs containing control Ab (10 µg/ml) or sVEGFR1 Ab (10 µg/ml) recovered from mice dorsum after the mice were infected or uninfected with SFTSV. ( C ) Representative cross-sections of H&E-staining of the Matrigel plugs, harvested as described in ( B ). Shown on the right are subset images of the regions outlined by rectangles on the left. Scale bars, 250 µm (left) and 50 µm (right), respectively. n = 3 biological replicates for each group. Statistical significance was determined by two-tailed unpaired Student t test. (Mock, *** P < 0.0001; SFTSV, *** P = 0.0006). ( D ) After the plugs were embedded in wax and sectioned, the sections were immune-stained with CD31 antibodies. Representative images of immunostaining are shown. Shown on the right are subset images of the regions outlined by rectangles on the left. Scale bars, 250 µm (left) and 50 µm (right), respectively. n = 3 biological replicates for each group. Statistical significance was determined by two-tailed unpaired Student t test. (Mock, *** P = 0.0007; SFTSV, ** P = 0.0039). Data information: Data shown are mean ± SD of three biological replicates. (** P < 0.01; *** P < 0.001). .

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Injection, Control, Infection, Staining, Two Tailed Test, Immunostaining

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: ( A ) PCR (left) and q-PCR (right) were used to detect the genotype and KO efficiency, respectively ( n = 3 biologically independent WT mice and KO mice). ND, Not Detected. ( B ) Experimental scheme of the SFTSV-infected mouse model. WT and EGR1-KO mice were infected with SFTSV at 1 × 10 5 TCID 50 , respectively. Liver, spleen, lung and kidney were isolated from two groups for analysis. ( C , D ) The percent of body weight and survival changes were monitored for 7 days. n = 5 biological replicates for each group. Statistical significance was determined by Log-rank test. (( D ): **, P = 0.0048) ( E ) sVEGFR1 (left) and SFTSV viral RNA mRNA (right) in blood cells from the WT and KO mice ( n = 5) were determined by qPCR. n = 5 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (sVEGFR1: ** P = 0.0087; SFTSV: * P = 0.0481) ( F ) The serum level of sVEGFR1 protein was measured by capture ELISA. Each point represents an individual. n = 5 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (* P = 0.0193). ( G ) Histopathological examinations of the tissues collected from the WT and KO mice. Tissues were collected from mice on day 4. The representative photographs of H&E were shown. The black arrows indicate hyperemia. Bars, 100 μm. ( H ) The infected mice were euthanized, and spleens were collected. sVEGFR1 mRNA in the spleens from the WT and KO mice were determined by qPCR. n = 5 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (* P = 0.0235). ( I ) sVEGFR1 and the viral NP protein in spleens were measured by western blotting (left). The sVEGFR1 and SFTSV-NP protein expression relative to GAPDH was shown (right), respectively. n = 3 biological replicates for Mock; n = 5 biological replicates for SFTSV. Statistical significance was determined by two-tailed unpaired Student t test. (sVEGFR1: ** P = 0.0054; NP: ** P = 0.0018). ( J ) Detection of sVEGFR1 and viral Gn protein by immunofluorescence in the spleens after SFTSV infection (left). SFTSV-Gn (green), sVEGFR1 (red) and nuclei (DAPI, blue) were visualized using either antibodies specific for Gn or sVEGFR1, or DAPI for nuclei. A merged image of green, red and blue channels is shown (merge). The quantifications of relative SFTSV-Gn and sVEGFR1 intensities were determined by ImageJ software (right). Scale bars, 25 µm. n = 5 biological replicates. Statistical significance was determined by two-tailed unpaired Student t test. (Gn: *** P < 0.0001; sVEGFR1: *** P < 0.0001). Data information: Data shown are mean ± SD of at least three biological replicates with each data point representing a biological experiment. (* P < 0.05; ** P < 0.01; *** P < 0.001). .

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Infection, Isolation, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Immunofluorescence, Software

Journal: EMBO Reports

Article Title: sVEGFR1 up-regulation via EGR1 impairs vascular repair in SFTSV-induced hemorrhage

doi: 10.1038/s44319-025-00541-2

Figure Lengend Snippet: The mechanism of hemorrhage that SFTSV-induced sVEGFR1 release through upregulation of EGR1 contributes to hemorrhage, suggesting that the EGR1/sVEGFR1/hemorrhage axis may be an effective target for the development of novel anti-hemorrhagic therapies during SFTSV infection.

Article Snippet: Mouse sVEGFR1 ELISA Kit , BOSTER , Cat#: EK0589.

Techniques: Infection