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mouse vegf quantikine elisa kit  (R&D Systems)


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    R&D Systems mouse vegf quantikine elisa kit
    T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by <t>ELISA.</t> n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures
    Mouse Vegf Quantikine Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 711 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+quantikine+elisa+kit/Mouse+VEGF+Quantikine+ELISA+Kit/pmc13625450-95-40-47
    Average 96 stars, based on 711 article reviews
    mouse vegf quantikine elisa kit - by Bioz Stars, 2026-11
    96/100 stars

    Images

    1) Product Images from "The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity"

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    Journal: Molecular Medicine

    doi: 10.1186/s10020-026-01651-w

    T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by ELISA. n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures
    Figure Legend Snippet: T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by ELISA. n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures

    Techniques Used: Migration, Activity Assay, Isolation, Cell Culture, Analysis, Counting, Co-culture Assay, Derivative Assay, WST Assay, Comparison, Enzyme-linked Immunosorbent Assay, Knock-Out

    Regulation of cytokines and angiogenesis markers in the tumor. A OLINK proteomic analysis of inflammatory cytokines and chemokines in the tumors of wildtype and IKKε KO mice at the end of the experiment. Heat map and Volcano Plot of significantly different regulations. n = 4/group ( B ) Cxcl1 and Cxcl2 ELISA. Cxcl1: WT n = 8, IKKε-KO n = 6; Cxcl2: WT n = 6, IKKε-KO n = 5. C Determination of CD31-positive cells as vessel markers in the tumors. Left panel: FACS analysis, n = 6 per group. Right panel: immunofluorescence images and quantitative analysis, n = 4–5 tumors per group; 3–5 areas per tumor were evaluated. CD31-positive areas were normalized to wildtype controls to facilitate comparison. D MMP-9 expression as assessed by Western blot and zymography, n = 5–6 per group. E HIF-1α expression as determined by immunofluorescence, the pictures show on representative stain from 6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. F VEGF expression analyzed by ELISA and Western Blot, n = 3/group. Student´s t-test, * P < 0.05, ** P < 0.01
    Figure Legend Snippet: Regulation of cytokines and angiogenesis markers in the tumor. A OLINK proteomic analysis of inflammatory cytokines and chemokines in the tumors of wildtype and IKKε KO mice at the end of the experiment. Heat map and Volcano Plot of significantly different regulations. n = 4/group ( B ) Cxcl1 and Cxcl2 ELISA. Cxcl1: WT n = 8, IKKε-KO n = 6; Cxcl2: WT n = 6, IKKε-KO n = 5. C Determination of CD31-positive cells as vessel markers in the tumors. Left panel: FACS analysis, n = 6 per group. Right panel: immunofluorescence images and quantitative analysis, n = 4–5 tumors per group; 3–5 areas per tumor were evaluated. CD31-positive areas were normalized to wildtype controls to facilitate comparison. D MMP-9 expression as assessed by Western blot and zymography, n = 5–6 per group. E HIF-1α expression as determined by immunofluorescence, the pictures show on representative stain from 6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. F VEGF expression analyzed by ELISA and Western Blot, n = 3/group. Student´s t-test, * P < 0.05, ** P < 0.01

    Techniques Used: Analysis, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Comparison, Expressing, Western Blot, Zymography, Staining

    Early regulation of cytokines and angiogenesis markers in the tumor. A Cxcl1 ELISA, ( B ) MMP-9 expression (Western Blot and zymography), ( C ) HIF-1α expression as determined by immunofluorescence. The pictures show a representative stain from 4–6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. D FACS analysis of CD31. + -cells as vessel markers in the tumors, ( n = 6/group). E VEGF expression analyzed by ELISA and Western Blot, ( F ) p65 expression and activity in nuclear extracts as determined by Western Blot and transcription factor ELISA. n = 3–5/group. Student´s t-test, * P < 0.05, ** P < 0.01
    Figure Legend Snippet: Early regulation of cytokines and angiogenesis markers in the tumor. A Cxcl1 ELISA, ( B ) MMP-9 expression (Western Blot and zymography), ( C ) HIF-1α expression as determined by immunofluorescence. The pictures show a representative stain from 4–6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. D FACS analysis of CD31. + -cells as vessel markers in the tumors, ( n = 6/group). E VEGF expression analyzed by ELISA and Western Blot, ( F ) p65 expression and activity in nuclear extracts as determined by Western Blot and transcription factor ELISA. n = 3–5/group. Student´s t-test, * P < 0.05, ** P < 0.01

    Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Zymography, Immunofluorescence, Staining, Analysis, Activity Assay

    Angiogenesis assay in HUVEC with and without depletion of IKKε by siRNA. A IKKε mRNA and protein expression in HUVEC cells after treatment with IKKε siRNA or a scrambled negative control as determined by PCR and Western Blot analysis, PCR: n = 3–4/group, Western Blot ( n = 2/group) ( B ) Tube formation assay, left side: microscopic pictures, right side: analysis of the total mesh area of naïve HUVEC, or HUVEC treated with NC and IKKε siRNA. n = 6–7/group. The scale bars represent 100 µm. C Spheroid sprout assay, left side: microscopic pictures, right side: analysis of cumulative sprout length (CSL) in spheroids of naïve HUVEC ( n = 15 spheroids), HUVEC treated with VEGF ( n = 38 spheroids) HUVEC-NC ( n = 25 spheroids) and HUVEC-IKKε siRNA ( n = 38 spheroids). Data were generated in 5 independent biological experiments. The scale bars represent 100 µm. One Way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001
    Figure Legend Snippet: Angiogenesis assay in HUVEC with and without depletion of IKKε by siRNA. A IKKε mRNA and protein expression in HUVEC cells after treatment with IKKε siRNA or a scrambled negative control as determined by PCR and Western Blot analysis, PCR: n = 3–4/group, Western Blot ( n = 2/group) ( B ) Tube formation assay, left side: microscopic pictures, right side: analysis of the total mesh area of naïve HUVEC, or HUVEC treated with NC and IKKε siRNA. n = 6–7/group. The scale bars represent 100 µm. C Spheroid sprout assay, left side: microscopic pictures, right side: analysis of cumulative sprout length (CSL) in spheroids of naïve HUVEC ( n = 15 spheroids), HUVEC treated with VEGF ( n = 38 spheroids) HUVEC-NC ( n = 25 spheroids) and HUVEC-IKKε siRNA ( n = 38 spheroids). Data were generated in 5 independent biological experiments. The scale bars represent 100 µm. One Way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

    Techniques Used: Angiogenesis Assay, Expressing, Negative Control, Western Blot, Analysis, Tube Formation Assay, Spheroids, Generated

    Related Articles

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    Article Snippet: The facultative intracellular pathogen Brucella spp. relies on the translocation of secreted effector proteins, such as BspH, to establish and maintain its persistent niche within mammalian and human host cell.. Following the cloning, expression, and purification of secreted protein BspH, we assessed its immunocompetence.. The recombinant protein (rBspH) was used to stimulate murine RAW 264.7 macrophages, and the production of cytokines along with hydrogen (H), calcium (Ca2+), and hydrogen peroxide (H2O2) ions was measured.

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    Article Snippet: .. A Mouse Quantikine ELISA Kit (R&D Systems) was used to quantify IL-10, TGF-β, IFN-γ, and TNFα levels in the conditioned media of the treated splenocytes. ..

    Article Title: PA0833 is a novel virulence factor that contributes to the pathogenicity of Pseudomonas aeruginosa lung infection.
    Article Snippet: Pseudomonas aeruginosa (P. aeruginosa), a major cause of severe respiratory infections, particularly ventilatorassociated pneumonia (VAP), employs various virulence mechanisms that remain incompletely characterized.. In this study, we report that PA0833, an OmpA family outer membrane protein, acts as a novel virulence factor that is involved in the regulation of bacterial pathogenicity.. We demonstrate that intranasal administration of recombinant PA0833 alone induces significant pulmonary inflammation in mice, characterized by alveolar disruption, neutrophil infiltration, and elevated pro-inflammatory cytokine levels in BALF, independent of live bacteria.

    Article Title: Exoproteome of calorie-restricted humans identifies complement deactivation as an immunometabolic checkpoint reducing inflammaging.
    Article Snippet: The serum protein levels of C3 and C3a in humans (Hycult Biotech) and mice (C3, Abcam; C3a, Thermo Fisher Scientific) were assessed using ELISA in both sexes. .. For C3a neutralization experiments, serum IL-1β, IL-6, TNF and MCP-1 levels were analyzed by the Mouse Quantikine ELISA Kit from R&D Systems. ..

    Article Title: Targeting KEAP1-mediated IKKβ degradation strategy for colitis-associated colorectal carcinogenesis: The potential of xanthohumol.
    Article Snippet: In colitis-associated colorectal cancer (CAC), the NF-κB pathway, especially IKKβ, drives inflammation and cancer progression.. However, no IKKβ inhibitors have been approved due to compensatory mechanisms.. The challenge is to develop an anti-tumor agent that effectively targets IKKβ while overcoming these compensatory pathways.

    Article Title: Silencing hepatic PCSK9 via novel chimeric AAV8 mitigates the progression of atherosclerosis by inhibiting inflammation in ApoE −/− mice
    Article Snippet: .. We measured plasma mouse PCSK9 levels using a mouse Quantikine ELISA Kit (catalog number, MPC900; R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions. .. Mouse MIF serum levels were measured using an MIF ELISA Kit (catalog number, E0698m; EIAab, Wuhan, China).

    Neutralization:

    Article Title: Exoproteome of calorie-restricted humans identifies complement deactivation as an immunometabolic checkpoint reducing inflammaging.
    Article Snippet: The serum protein levels of C3 and C3a in humans (Hycult Biotech) and mice (C3, Abcam; C3a, Thermo Fisher Scientific) were assessed using ELISA in both sexes. .. For C3a neutralization experiments, serum IL-1β, IL-6, TNF and MCP-1 levels were analyzed by the Mouse Quantikine ELISA Kit from R&D Systems. ..

    Concentration Assay:

    Article Title: Brucella secreted effector BspH stimulates intracellular ion flux and confers anti-Brucella immunity.
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    Clinical Proteomics:

    Article Title: Silencing hepatic PCSK9 via novel chimeric AAV8 mitigates the progression of atherosclerosis by inhibiting inflammation in ApoE −/− mice
    Article Snippet: .. We measured plasma mouse PCSK9 levels using a mouse Quantikine ELISA Kit (catalog number, MPC900; R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions. .. Mouse MIF serum levels were measured using an MIF ELISA Kit (catalog number, E0698m; EIAab, Wuhan, China).



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    T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by <t>ELISA.</t> n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures
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    circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of <t>IL-10,</t> TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.
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    Image Search Results


    T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by ELISA. n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: T-cell migration and activity. A T-cell migration towards a FCS gradient or conditioned medium from B16BL6 melanoma cells. Pan T-cells were isolated from the spleens of wildtype (WT) and IKKε KO mice and plated into cell culture insert filled with T-cell medium containing 0.1% FCS. The bottom chamber was filled either with medium containing 0.1% FCS (neg ctrl), 10% FCS (pos ctrl) or conditioned medium from B16BL6 cells without FCS (B16). Cell migration of different T-cell subtypes to the lower channel was determined by FACS analysis. Cell numbers were normalized to counting beads and negative controls. n = 3–6. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01. B — D Effects of co-culture of B16BL6 melanoma cells with CD8⁺ and γδ-T-cells, respectively, derived from the spleens of wildtype (WT) and IKKε KO mice. B Cell proliferation determined by WST assay, B16BL6 monocultures were set as 1 for better comparison. One-way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, **P < 0.01 significant difference in comparison to B16BL6 monoculture ( C ) IFN-γ levels analyzed by cytokine bead array (CBA) and ( D ) granzyme B release determined by ELISA. n = 4–10/group. C and D The T-cell monocultures of wildtype and IKKε knock-out mice were set as 1 for better comparison. Student´s t-test, * P < 0.05, significant difference in comparison to wildtype, # P < 0.05, ## P < 0.01, ### P < 0.001 significant difference in comparison to T-cell monocultures

    Article Snippet: Granzyme B, Cxcl1, Cxcl2 and VEGF protein levels were determined from cell and tumor protein lysates using Mouse Granzyme B ELISA Kit (Invitrogen/Thermo Fisher Scientific, Dreieich, Germany #BMS6079), Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B), Mouse CXCL2/MIP-2 DuoSet ELISA (#DY452) and Mouse VEGF Quantikine ELISA Kit (#MMVOO-1) (all R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer’s instructions.

    Techniques: Migration, Activity Assay, Isolation, Cell Culture, Analysis, Counting, Co-culture Assay, Derivative Assay, WST Assay, Comparison, Enzyme-linked Immunosorbent Assay, Knock-Out

    Regulation of cytokines and angiogenesis markers in the tumor. A OLINK proteomic analysis of inflammatory cytokines and chemokines in the tumors of wildtype and IKKε KO mice at the end of the experiment. Heat map and Volcano Plot of significantly different regulations. n = 4/group ( B ) Cxcl1 and Cxcl2 ELISA. Cxcl1: WT n = 8, IKKε-KO n = 6; Cxcl2: WT n = 6, IKKε-KO n = 5. C Determination of CD31-positive cells as vessel markers in the tumors. Left panel: FACS analysis, n = 6 per group. Right panel: immunofluorescence images and quantitative analysis, n = 4–5 tumors per group; 3–5 areas per tumor were evaluated. CD31-positive areas were normalized to wildtype controls to facilitate comparison. D MMP-9 expression as assessed by Western blot and zymography, n = 5–6 per group. E HIF-1α expression as determined by immunofluorescence, the pictures show on representative stain from 6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. F VEGF expression analyzed by ELISA and Western Blot, n = 3/group. Student´s t-test, * P < 0.05, ** P < 0.01

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: Regulation of cytokines and angiogenesis markers in the tumor. A OLINK proteomic analysis of inflammatory cytokines and chemokines in the tumors of wildtype and IKKε KO mice at the end of the experiment. Heat map and Volcano Plot of significantly different regulations. n = 4/group ( B ) Cxcl1 and Cxcl2 ELISA. Cxcl1: WT n = 8, IKKε-KO n = 6; Cxcl2: WT n = 6, IKKε-KO n = 5. C Determination of CD31-positive cells as vessel markers in the tumors. Left panel: FACS analysis, n = 6 per group. Right panel: immunofluorescence images and quantitative analysis, n = 4–5 tumors per group; 3–5 areas per tumor were evaluated. CD31-positive areas were normalized to wildtype controls to facilitate comparison. D MMP-9 expression as assessed by Western blot and zymography, n = 5–6 per group. E HIF-1α expression as determined by immunofluorescence, the pictures show on representative stain from 6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. F VEGF expression analyzed by ELISA and Western Blot, n = 3/group. Student´s t-test, * P < 0.05, ** P < 0.01

    Article Snippet: Granzyme B, Cxcl1, Cxcl2 and VEGF protein levels were determined from cell and tumor protein lysates using Mouse Granzyme B ELISA Kit (Invitrogen/Thermo Fisher Scientific, Dreieich, Germany #BMS6079), Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B), Mouse CXCL2/MIP-2 DuoSet ELISA (#DY452) and Mouse VEGF Quantikine ELISA Kit (#MMVOO-1) (all R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer’s instructions.

    Techniques: Analysis, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Comparison, Expressing, Western Blot, Zymography, Staining

    Early regulation of cytokines and angiogenesis markers in the tumor. A Cxcl1 ELISA, ( B ) MMP-9 expression (Western Blot and zymography), ( C ) HIF-1α expression as determined by immunofluorescence. The pictures show a representative stain from 4–6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. D FACS analysis of CD31. + -cells as vessel markers in the tumors, ( n = 6/group). E VEGF expression analyzed by ELISA and Western Blot, ( F ) p65 expression and activity in nuclear extracts as determined by Western Blot and transcription factor ELISA. n = 3–5/group. Student´s t-test, * P < 0.05, ** P < 0.01

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: Early regulation of cytokines and angiogenesis markers in the tumor. A Cxcl1 ELISA, ( B ) MMP-9 expression (Western Blot and zymography), ( C ) HIF-1α expression as determined by immunofluorescence. The pictures show a representative stain from 4–6 independent analyses, the diagram shows the densitometric analysis of all experiments, scale bar represents 100 µm. D FACS analysis of CD31. + -cells as vessel markers in the tumors, ( n = 6/group). E VEGF expression analyzed by ELISA and Western Blot, ( F ) p65 expression and activity in nuclear extracts as determined by Western Blot and transcription factor ELISA. n = 3–5/group. Student´s t-test, * P < 0.05, ** P < 0.01

    Article Snippet: Granzyme B, Cxcl1, Cxcl2 and VEGF protein levels were determined from cell and tumor protein lysates using Mouse Granzyme B ELISA Kit (Invitrogen/Thermo Fisher Scientific, Dreieich, Germany #BMS6079), Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B), Mouse CXCL2/MIP-2 DuoSet ELISA (#DY452) and Mouse VEGF Quantikine ELISA Kit (#MMVOO-1) (all R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer’s instructions.

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Zymography, Immunofluorescence, Staining, Analysis, Activity Assay

    Angiogenesis assay in HUVEC with and without depletion of IKKε by siRNA. A IKKε mRNA and protein expression in HUVEC cells after treatment with IKKε siRNA or a scrambled negative control as determined by PCR and Western Blot analysis, PCR: n = 3–4/group, Western Blot ( n = 2/group) ( B ) Tube formation assay, left side: microscopic pictures, right side: analysis of the total mesh area of naïve HUVEC, or HUVEC treated with NC and IKKε siRNA. n = 6–7/group. The scale bars represent 100 µm. C Spheroid sprout assay, left side: microscopic pictures, right side: analysis of cumulative sprout length (CSL) in spheroids of naïve HUVEC ( n = 15 spheroids), HUVEC treated with VEGF ( n = 38 spheroids) HUVEC-NC ( n = 25 spheroids) and HUVEC-IKKε siRNA ( n = 38 spheroids). Data were generated in 5 independent biological experiments. The scale bars represent 100 µm. One Way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

    Journal: Molecular Medicine

    Article Title: The protein kinase IKK epsilon shapes the melanoma tumor microenvironment by modulating angiogenesis and the T-cell immunity

    doi: 10.1186/s10020-026-01651-w

    Figure Lengend Snippet: Angiogenesis assay in HUVEC with and without depletion of IKKε by siRNA. A IKKε mRNA and protein expression in HUVEC cells after treatment with IKKε siRNA or a scrambled negative control as determined by PCR and Western Blot analysis, PCR: n = 3–4/group, Western Blot ( n = 2/group) ( B ) Tube formation assay, left side: microscopic pictures, right side: analysis of the total mesh area of naïve HUVEC, or HUVEC treated with NC and IKKε siRNA. n = 6–7/group. The scale bars represent 100 µm. C Spheroid sprout assay, left side: microscopic pictures, right side: analysis of cumulative sprout length (CSL) in spheroids of naïve HUVEC ( n = 15 spheroids), HUVEC treated with VEGF ( n = 38 spheroids) HUVEC-NC ( n = 25 spheroids) and HUVEC-IKKε siRNA ( n = 38 spheroids). Data were generated in 5 independent biological experiments. The scale bars represent 100 µm. One Way ANOVA with Tukey´s multiple comparisons test, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

    Article Snippet: Granzyme B, Cxcl1, Cxcl2 and VEGF protein levels were determined from cell and tumor protein lysates using Mouse Granzyme B ELISA Kit (Invitrogen/Thermo Fisher Scientific, Dreieich, Germany #BMS6079), Mouse CXCL1/KC Quantikine ELISA Kit (#MKC00B), Mouse CXCL2/MIP-2 DuoSet ELISA (#DY452) and Mouse VEGF Quantikine ELISA Kit (#MMVOO-1) (all R&D Systems, Minneapolis, Minnesota, USA) according to the manufacturer’s instructions.

    Techniques: Angiogenesis Assay, Expressing, Negative Control, Western Blot, Analysis, Tube Formation Assay, Spheroids, Generated

    circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

    Journal: Non-coding RNA Research

    Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

    doi: 10.1016/j.ncrna.2026.03.003

    Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

    Article Snippet: For mouse experiments, mouse IL-10 was measured using the Mouse IL-10 ELISA Kit (R&D Systems, Cat# M1000B), and mouse TGF-β1 was measured using the Mouse TGF beta-1 ELISA Kit (Invitrogen, Cat# BMS608-4), following the manufacturers’ instructions.

    Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration