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platelet derived growth factor pdgf bb  (R&D Systems)


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    R&D Systems platelet derived growth factor pdgf bb
    Platelet Derived Growth Factor Pdgf Bb, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Fig. 4 Validation of the RNAseq data via mRNA and protein expression employing RT-qPCR, <t>ELISA</t> and CBA. The DEGs were further validated by RT-qPCR and analysis of protein expression. Four differentially regulated genes per cell line are depicted. Gene expression was normalized to ACTB (housekeeping gene) and the corresponding control vector cells (2−ΔΔct method). A qPCR analysis of four representative DEGs from HCT-116 5-LO KO cells. B Secretion of TGFβ2 into the cell culture supernatants of HCT-116 5-LO KO cells measured via ELISA. Depicted are the relative TGFβ2 amounts in % compared to the vector control (mean TGFβ2 secretion from vector control cells: 231.9 ± 140 pg/ µg total protein). C qPCR analysis of four representative DEGs from HT-29 5-LO KO cells and D U-2 OS cells. E Cytokine release from U-2 OS 5-LO KO cells. Data are depicted as relative cytokine amounts in % compared to the corresponding control vector cells. PDGF-AA was assessed by ELISA while fractalkine (CX3CL1) and MCP-1 (CCL2) were measured via FACS employing a cytometric bead array (mean secretion from vector control cells: fractalkine, 64.9 ± 9.2 pg/µg protein; MCP-1, 978.2 ± 101.8 pg/µg protein; PDGF-AA, 255,6 ± 95.6 pg/µg protein). The complete mRNA expression data can be found in supplementary Fig. 4 and supplementary table 1. All data are presented as mean + SD of three independent experiments. Asterisks indicate significant changes vs. control vector cells. *P ≤0.05, **P ≤0.01, ***P ≤0.001.
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    Fig. 4 Validation of the RNAseq data via mRNA and protein expression employing RT-qPCR, <t>ELISA</t> and CBA. The DEGs were further validated by RT-qPCR and analysis of protein expression. Four differentially regulated genes per cell line are depicted. Gene expression was normalized to ACTB (housekeeping gene) and the corresponding control vector cells (2−ΔΔct method). A qPCR analysis of four representative DEGs from HCT-116 5-LO KO cells. B Secretion of TGFβ2 into the cell culture supernatants of HCT-116 5-LO KO cells measured via ELISA. Depicted are the relative TGFβ2 amounts in % compared to the vector control (mean TGFβ2 secretion from vector control cells: 231.9 ± 140 pg/ µg total protein). C qPCR analysis of four representative DEGs from HT-29 5-LO KO cells and D U-2 OS cells. E Cytokine release from U-2 OS 5-LO KO cells. Data are depicted as relative cytokine amounts in % compared to the corresponding control vector cells. PDGF-AA was assessed by ELISA while fractalkine (CX3CL1) and MCP-1 (CCL2) were measured via FACS employing a cytometric bead array (mean secretion from vector control cells: fractalkine, 64.9 ± 9.2 pg/µg protein; MCP-1, 978.2 ± 101.8 pg/µg protein; PDGF-AA, 255,6 ± 95.6 pg/µg protein). The complete mRNA expression data can be found in supplementary Fig. 4 and supplementary table 1. All data are presented as mean + SD of three independent experiments. Asterisks indicate significant changes vs. control vector cells. *P ≤0.05, **P ≤0.01, ***P ≤0.001.
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    Fig. 4 Validation of the RNAseq data via mRNA and protein expression employing RT-qPCR, <t>ELISA</t> and CBA. The DEGs were further validated by RT-qPCR and analysis of protein expression. Four differentially regulated genes per cell line are depicted. Gene expression was normalized to ACTB (housekeeping gene) and the corresponding control vector cells (2−ΔΔct method). A qPCR analysis of four representative DEGs from HCT-116 5-LO KO cells. B Secretion of TGFβ2 into the cell culture supernatants of HCT-116 5-LO KO cells measured via ELISA. Depicted are the relative TGFβ2 amounts in % compared to the vector control (mean TGFβ2 secretion from vector control cells: 231.9 ± 140 pg/ µg total protein). C qPCR analysis of four representative DEGs from HT-29 5-LO KO cells and D U-2 OS cells. E Cytokine release from U-2 OS 5-LO KO cells. Data are depicted as relative cytokine amounts in % compared to the corresponding control vector cells. PDGF-AA was assessed by ELISA while fractalkine (CX3CL1) and MCP-1 (CCL2) were measured via FACS employing a cytometric bead array (mean secretion from vector control cells: fractalkine, 64.9 ± 9.2 pg/µg protein; MCP-1, 978.2 ± 101.8 pg/µg protein; PDGF-AA, 255,6 ± 95.6 pg/µg protein). The complete mRNA expression data can be found in supplementary Fig. 4 and supplementary table 1. All data are presented as mean + SD of three independent experiments. Asterisks indicate significant changes vs. control vector cells. *P ≤0.05, **P ≤0.01, ***P ≤0.001.
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    Fig. 4 Validation of the RNAseq data via mRNA and protein expression employing RT-qPCR, ELISA and CBA. The DEGs were further validated by RT-qPCR and analysis of protein expression. Four differentially regulated genes per cell line are depicted. Gene expression was normalized to ACTB (housekeeping gene) and the corresponding control vector cells (2−ΔΔct method). A qPCR analysis of four representative DEGs from HCT-116 5-LO KO cells. B Secretion of TGFβ2 into the cell culture supernatants of HCT-116 5-LO KO cells measured via ELISA. Depicted are the relative TGFβ2 amounts in % compared to the vector control (mean TGFβ2 secretion from vector control cells: 231.9 ± 140 pg/ µg total protein). C qPCR analysis of four representative DEGs from HT-29 5-LO KO cells and D U-2 OS cells. E Cytokine release from U-2 OS 5-LO KO cells. Data are depicted as relative cytokine amounts in % compared to the corresponding control vector cells. PDGF-AA was assessed by ELISA while fractalkine (CX3CL1) and MCP-1 (CCL2) were measured via FACS employing a cytometric bead array (mean secretion from vector control cells: fractalkine, 64.9 ± 9.2 pg/µg protein; MCP-1, 978.2 ± 101.8 pg/µg protein; PDGF-AA, 255,6 ± 95.6 pg/µg protein). The complete mRNA expression data can be found in supplementary Fig. 4 and supplementary table 1. All data are presented as mean + SD of three independent experiments. Asterisks indicate significant changes vs. control vector cells. *P ≤0.05, **P ≤0.01, ***P ≤0.001.

    Journal: Cancer gene therapy

    Article Title: Knock-out of 5-lipoxygenase in overexpressing tumor cells-consequences on gene expression and cellular function.

    doi: 10.1038/s41417-022-00531-9

    Figure Lengend Snippet: Fig. 4 Validation of the RNAseq data via mRNA and protein expression employing RT-qPCR, ELISA and CBA. The DEGs were further validated by RT-qPCR and analysis of protein expression. Four differentially regulated genes per cell line are depicted. Gene expression was normalized to ACTB (housekeeping gene) and the corresponding control vector cells (2−ΔΔct method). A qPCR analysis of four representative DEGs from HCT-116 5-LO KO cells. B Secretion of TGFβ2 into the cell culture supernatants of HCT-116 5-LO KO cells measured via ELISA. Depicted are the relative TGFβ2 amounts in % compared to the vector control (mean TGFβ2 secretion from vector control cells: 231.9 ± 140 pg/ µg total protein). C qPCR analysis of four representative DEGs from HT-29 5-LO KO cells and D U-2 OS cells. E Cytokine release from U-2 OS 5-LO KO cells. Data are depicted as relative cytokine amounts in % compared to the corresponding control vector cells. PDGF-AA was assessed by ELISA while fractalkine (CX3CL1) and MCP-1 (CCL2) were measured via FACS employing a cytometric bead array (mean secretion from vector control cells: fractalkine, 64.9 ± 9.2 pg/µg protein; MCP-1, 978.2 ± 101.8 pg/µg protein; PDGF-AA, 255,6 ± 95.6 pg/µg protein). The complete mRNA expression data can be found in supplementary Fig. 4 and supplementary table 1. All data are presented as mean + SD of three independent experiments. Asterisks indicate significant changes vs. control vector cells. *P ≤0.05, **P ≤0.01, ***P ≤0.001.

    Article Snippet: PDGF-AA was measured with the DuoSet® Human PDGF-AA ELISA Kit (R&D systems) following the manufacturer’s protocol.

    Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Gene Expression, Control, Plasmid Preparation, Cell Culture