human vegf vegfa elisa kit  (Boster Bio)

Journal: Tumour Virus Research

Article Title: PRMT5–mediated symmetric dimethylation of SHBs at Arg169 stabilizes SHBs and promotes angiogenesis and tumor growth

doi: 10.1016/j.tvr.2026.200340

Figure Lengend Snippet: Arg169 symmetric dimethylation is required for SHBs–driven angiogenesis and tumor growth. (A) WB analysis of SHBs and BIP expression in stably transduced Huh7 and HepG2 cells (Vector, SHBs, and SHBs/R169K). (B) ELISA measurement of VEGFA levels in the supernatants of Huh7/HepG2–Vector, Huh7/HepG2–SHBs, or Huh7/HepG2–SHBs/R169K cells. (C) Endothelial tube formation assay. EA.hy926 cells were cultured with conditioned media (CM) from Huh7 or HepG2 stable lines (Vector, SHBs, SHBs/R169K). Representative images and quantification of mesh numbers are shown. (D) Transwell migration assay. EA.hy926 cells were assessed for migration in response to CM from the indicated stable lines. Representative images and quantification of migrated cell numbers per field are shown. (E) Representative images of excised subcutaneous xenograft tumors derived from Huh7–Vector, Huh7–SHBs, or Huh7–SHBs/R169K cells. (F) Tumor growth curves (tumor volume over time) for the indicated xenograft groups. (G) Tumor weights at endpoint. (H) Representative immunohistochemical staining of xenograft tumors for CD31 and SHBs, with quantification of microvessel density (MVD) based on CD31 staining. Data are presented as mean ± SD; ∗ P < 0.05 as indicated.

Article Snippet: The supernatants were collected from cells, and VEGFA was quantified by using the Human VEGF/VEGFA ELISA Kit (Boster, # EK0539).

Techniques: Expressing, Stable Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Endothelial Tube Formation Assay, Cell Culture, Transwell Migration Assay, Migration, Derivative Assay, Immunohistochemical staining, Staining

Journal: Ophthalmology Science

Article Title: Serial Aqueous Humor Proteomics in Diabetic Macular Edema

doi: 10.1016/j.xops.2025.101056

Figure Lengend Snippet: Abundance of proteins significantly associated with incomplete response to anti-VEGF therapy. A total of 10 proteins were found to be significantly regulated in substantial edema versus no edema. A, Upregulated proteins determined by mass spectrometry at time points 1 to 3 (T1-T3) included afamin (AFM), alpha-1B-glycoprotein (A1BG), and angiotensinogen (AGT). B, Downregulated proteins included cystatin-C (CST3), clusterin (CLU), apolipoprotein E (APOE), epidermal growth factor-containing fibulin-like extracellular matrix protein (EFEMP1), Ig lambda chain V-I (IGLV1), latent transforming growth factor beta-binding protein 2 (LTBP2), and reelin (RELN). C, The increased level of afamin was confirmed by ELISA. D, VEGFA concentration assessed by ELISA. There was no significant difference in VEGFA between the substantial edema and no edema groups. Box plots indicate median, min and max and first and third quartiles. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001. ELISA = enzyme-linked immunosorbent assay; LFQ = Label-Free Quantification.

Article Snippet: Samples were diluted 500-fold for afamin (ELH-AFM-1, RayBiotech) and 2-fold for VEGFA (DVE00, R&D Systems).

Techniques: Mass Spectrometry, Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Quantitative Proteomics

Journal: Advanced Science

Article Title: Intercellular Horizontal Transfer of TXNDC5 mRNA via Extracellular Vesicles Contributes to Tumor‐Associated Macrophage‐Mediated Prostate Cancer Metastasis

doi: 10.1002/advs.202511052

Figure Lengend Snippet: Primary lesions of mPCa exhibit increased M2 macrophage infiltration and a higher M2‐EVs/Ti‐EVs ratio compared to those of nmPCa. (A) Schematic diagram of the technical workflow to characterize M2 macrophages and M2 EVs in the primary lesions of PCa. (B) UMAP projection of all macrophages from the integrated scRNA‐seq dataset, colored by annotated cell subtypes: M1 macrophages, M2 macrophages, and Mixed macrophages. Cell numbers for each subset were indicated. (C) Analysis of the relative proportion of macrophage groups based on the integrated scRNA‐seq dataset in nmPCa and mPCa. (D) Association between the M2 phenotype and clinical outcome. M2 phenotype was defined by the expression of CD68 , CD163 , CD206 , IL10 , ARG1 , TGFB1 , VEGFA , and CCL22 . (E–G) Expression of M2 macrophage‐associated markers (represented by CD68, CD163, and CD206) in primary mPCa and nmPCa sites was shown by IHC (E, F) and IF(G). LR: low risk (patients with PSA value below 10 ng/mL, Gleason score below or equal to 7, and cT1‐cT2a disease); HR: high risk (patients with PSA value above 20 ng/mL, Gleason score above 7, cT2c‐cT4 disease, or a node‐positive disease). Scale bar, 50 µm. (H) Representative TEM images of Ti‐EVs from primary mPCa and nmPCa sites. Scale bar, 200 nm. (I) Size distribution of Ti‐EVs from primary mPCa and nmPCa sites showed by nFCM. (J) Western blotting of EV marker proteins (CD63, ALIX, and CD9) and contaminating protein (GM130). CL: cell lysate of tissue. (K, L) The proportions of CD68 + CD206 + EVs measured with nFCM in total Ti‐EVs from mPCa ( n = 5) compared to those from nmPCa ( n = 5). All experiments were repeated three times. Data presented as the mean ± SD. * , p < 0.05; ** , p < 0.01; *** , p < 0.001; **** , p < 0.0001; and ns for non‐significant data. Φ: macrophages.

Article Snippet: For the measurement of secretory protein concentrations, conditioned medium was collected and centrifuged for 20 min at 10 000 × g, and the supernatant was used to test the concentrations of TGF‐β1 (E‐EL‐0162, Elabscience), CCL22 (E‐EL‐H0029, Elabscience), and VEGFA (E‐EL‐H0111, Elabscience) with ELISA kits according to the manufacturer's instructions.

Techniques: Expressing, Western Blot, Marker

Journal: Advanced Science

Article Title: Intercellular Horizontal Transfer of TXNDC5 mRNA via Extracellular Vesicles Contributes to Tumor‐Associated Macrophage‐Mediated Prostate Cancer Metastasis

doi: 10.1002/advs.202511052

Figure Lengend Snippet: CCM of M2 macrophages promotes migration and invasion of PCa cells. (A) Illustration of the strategy used to induce M0 and M2 macrophages in human leukemia monocytic THP‐1 cells. THP‐1 cells were differentiated into M0 macrophages by incubation with 100 ng/mL phorbol‐12‐myristate‐13‐acetate (PMA) for 48 h. M0 macrophages were polarized into M2 macrophages by culturing in 20 ng/mL IL‐4 and IL‐10 for 48 h. (B) Characterization of morphological changes in the course of differentiation from THP‐1 cells to M2 macrophages under a light microscope. Scale bars, 200 µm (100×), 100 µm (200×), 50 µm (400×). (C) ELISA revealed elevated levels of secretory TGF‐β, CCL22, and VEGFA in the CCM of M2 macrophages compared with those of M0 macrophages. (D) Evaluation of M2 macrophage‐associated protein markers by flow cytometry before and after differentiation. (E) Verification of classical M2‐associated genes by qRT‐PCR in M0 and M2 macrophages. Gene expression normalized to GAPDH . (F) The proportional change of CD68 + CD163 + cells upon induction was shown by IF. Scale bar, 100 µm. (G, H) Migration and invasion assays in M2 CCM‐treated versus M0 CCM‐treated DU145 (G) and PC3 (H) cells. (I, J) The wound healing assay showed different migration rates of DU145 (I) and PC3 (J) cells upon M2 CCM treatment. All experiments were repeated three times. Data presented as the mean ± SD. * , p < 0.05; ** , p < 0.01; *** , p < 0.001; **** , p < 0.0001; and ns for non‐significant data.

Article Snippet: For the measurement of secretory protein concentrations, conditioned medium was collected and centrifuged for 20 min at 10 000 × g, and the supernatant was used to test the concentrations of TGF‐β1 (E‐EL‐0162, Elabscience), CCL22 (E‐EL‐H0029, Elabscience), and VEGFA (E‐EL‐H0111, Elabscience) with ELISA kits according to the manufacturer's instructions.

Techniques: Migration, Incubation, Light Microscopy, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Quantitative RT-PCR, Gene Expression, Wound Healing Assay

Journal: Cancer Cell International

Article Title: Heme oxygenase-1 leads to cisplatin resistance in nasopharyngeal carcinoma by reducing oxidative stress and ferroptosis

doi: 10.1186/s12935-025-03908-6

Figure Lengend Snippet: HO-1 contributes to CDDP resistance in NPC cells. Parental HK1 and C666-1 cells were administered a HO-1 inducer Hemin while HK1/R and C666-1/R cells were administered a HO-1 inhibitor ZnPP. A - B : CCK-8 assay for viability of cells and the IC50 values of CDDP. The cells were subjected to 5 µM CDDP treatment. C : Colony formation assays for NPC cell growth. D : Scratch tests for NPC cell migration. E : Transwell assays for NPC cell invasion. F : TUNEL assays for cell apoptosis. G : Immunofluorescence staining for γ-H2AX expression. H , DCFH-DA staining for ROS production. The conditioned media of these cell cultures were collected for HUVEC incubation. H : Angiogenesis ability of the HUVECs. I , ELISA tests for VEGFA and MMP9 levels in conditioned media. Six independent experiments were performed. Data are present as dot and bars, with each dot indicating one independent experiment. Differences were compared by the ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: The contents of malondialdehyde (MDA), superoxide dismutase (SOD), vascular endothelial growth factor A (VEGFA), and matrix metalloproteinase-9 (MMP9) were analyzed using MDA assay kit (D799761, Sangon Biotech Co., Ltd., Shanghai, China), SOD activity assay kit (D799593, Sangon), Human VEGFA enzyme-linked immunosorbent assay (ELISA) (E-EL-H0111, Elabscience Biotechnology Co., Ltd., Wuhai, Hubei, China), and Human MMP-9 ELISA Kit (E-EL-H6075, Elabscience).

Techniques: CCK-8 Assay, Migration, TUNEL Assay, Immunofluorescence, Staining, Expressing, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Cancer Discovery

Article Title: Requirement for Cyclin D1 Underlies Cell-Autonomous HIF2 Dependence in Kidney Cancer

doi: 10.1158/2159-8290.CD-24-1378

Figure Lengend Snippet: Failure to downregulate VEGFA and CCND1 confers resistance to PT2399 in vivo . A, VEGFA levels from conditioned media of 786-O cells that stably express firefly luciferase, dCas9-VP64, and the indicated CRISPRa sgRNAs after treatment with 2 μmol/L PT2399 or DMSO for 48 hours. The VEGF levels were normalized to the NT-a2 expressing cells treated with DMSO. Data are the means ± SD of n = 3 biological replicates. **, P < 0.01, and NS, unpaired t test. B, Cellular proliferation assays of 786-O cells that stably express firefly luciferase, dCas9-VP64, CRISPRa sgRNA VEGFA-a1, and either cyclin D1 (WT) or the EV. The cells were treated with 2 μmol/L PT2399 or DMSO for 16 days. Data are the means ± SD of n = 3 biological replicates and were normalized to the DMSO-treated cells for the respective cell lines (EV or WT). *, P < 0.05; ***, P < 0.001; unpaired t test. C, Immunoblot analysis of orthotopic xenografts formed by cells from ( B ) that were treated with PT2399 (30 mg/kg) or vehicle daily for 5 days by oral gavage. D, Average relative BLI intensity over time of orthotopic xenografts formed by cells from ( B ) that were treated with PT2399 (30 mg/kg) or vehicle daily for 28 days by oral gavage. For each mouse, BLI readings were normalized to the BLI value from the day the treatment was started. Data are the mean ± SEM from three independent experiments. ***, P < 0.001, and NS, two-way ANOVA. E and F, Kaplan–Meier survival curves for mice in D . Rx bar indicates length of treatment. Log-rank (Mantel–Cox) test, with indicated P values. NS, not significant.

Article Snippet: VEGF levels in conditioned media of cells treated with PT2399 or DMSO was measured in triplicate using Human VEGFA Quantikine ELISA Kit (R&D Systems, DVE00) according to the manufacturer’s instructions.

Techniques: In Vivo, Stable Transfection, Luciferase, Expressing, Western Blot