human pd 1 quantikine elisa  (R&D Systems)

Journal: Hepatology Communications

Article Title: Gut Dysbiosis and Fecal Calprotectin Predict Response to Immune Checkpoint Inhibitors in Patients With Hepatocellular Carcinoma

doi: 10.1002/hep4.1905

Figure Lengend Snippet: Changes in markers of gut dysbiosis (AE ratio), intestinal inflammation, and permeability (calprotectin, zonulin‐1, LBP) and PD‐L1 serum levels over the treatment period. Abbreviations: AE, Akkermansia to Enterobacteriaceae ratio; LBP, lipopolysaccharide binding protein; PD‐L1, programmed death ligand 1.

Article Snippet: Gut microbiota analysis and quantification of fecal calprotectin, lipopolysaccharide binding protein (LBP), and zonulin‐1 serum levels were performed at the above time points, as reported. ( , ) Commercial enzyme‐linked immunosorbent assays (ELISA) for the detection of serum levels of PD‐L1 (#DB7H10; R&D Systems Inc., Minneapolis, MN) and LBP (human LBP; Hycult Biotech, the Netherlands) were also performed.

Techniques: Permeability, Binding Assay

Journal: International Journal of Nephrology

Article Title: Bevacizumab Increases Endothelin-1 Production via Forkhead Box Protein O1 in Human Glomerular Microvascular Endothelial Cells In Vitro

doi: 10.1155/2021/8381115

Figure Lengend Snippet: Bevacizumab induced ET-1 production in cultured human glomerular microvascular endothelial cells (hGECs). hGECs were plated in 24-well plates (1 × 10 5 cells/well) for ELISA or 60 mm culture dishes (1 × 10 6 cells/dish) for RT-PCR. hGECs that were 90% confluent were serum-starved for 24 h before the experiments were performed. hGECs were treated with 0.1 or 1 µ M bevacizumab for 8 h, RNA was extracted from the cells, and ET-1 mRNA level was examined by real-time RT-PCR (a). The medium was collected and ET-1 protein level was examined by the ELISA (b). Data shown are means ± SD ( n = 3; ∗ p < 0.05). The experiments were repeated at least three times, with reproducible results.

Article Snippet: The concentration of ET-1 in the supernatant was determined by sandwich enzyme-linked immunosorbent assay (ELISA) using a human ET-1 Quantikine ELISA kit (R&D Systems, Abingdon, UK).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: International Journal of Nephrology

Article Title: Bevacizumab Increases Endothelin-1 Production via Forkhead Box Protein O1 in Human Glomerular Microvascular Endothelial Cells In Vitro

doi: 10.1155/2021/8381115

Figure Lengend Snippet: The Akt pathway was inactivated in human glomerular microvascular endothelial cells (hGECs) treated with bevacizumab. hGECs were plated in 24-well plates (1 × 10 5 cells/well) for ELISA or 60 mm culture dishes (1 × 10 6 cells/dish) for RT-PCR and western blot. hGECs that were 90% confluent were serum-starved for 24 h before the experiments were performed. hGECs were treated with 1 µ M bevacizumab or 10 µ M LY294002 (PI3K/Akt inhibitor) for 8 h, and western blots were used to evaluate the changes in Akt phosphorylation after bevacizumab treatment (a). The levels of ET-1 mRNA (b) and ET-1 protein (c) were measured using real-time RT-PCR and the ELISA after LY294002 treatment. Data shown are means ± SD ( n = 3; ∗ p < 0.05). The experiments were repeated at least three times, with reproducible results.

Article Snippet: The concentration of ET-1 in the supernatant was determined by sandwich enzyme-linked immunosorbent assay (ELISA) using a human ET-1 Quantikine ELISA kit (R&D Systems, Abingdon, UK).

Techniques: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Phospho-proteomics, Quantitative RT-PCR

Journal: International Journal of Nephrology

Article Title: Bevacizumab Increases Endothelin-1 Production via Forkhead Box Protein O1 in Human Glomerular Microvascular Endothelial Cells In Vitro

doi: 10.1155/2021/8381115

Figure Lengend Snippet: The nuclear localization of FoxO1 was increased in human glomerular microvascular endothelial cells (hGECs) treated with bevacizumab. hGECs were plated in 24-well plates (1 × 10 5 cells/well) for ELISA or 60 mm culture dishes (1 × 10 6 cells/dish) for RT-PCR and western blot. hGECs that were 90% confluent were serum-starved for 24 h before the experiments were performed. hGECs were treated with 1 µ M bevacizumab and 0.1 µ M AS1842856 (FoxO1 inhibitor) for 8 h, and western blots were used to evaluate the changes in FoxO1 protein level in cytosolic (cyto) and nuclear (nu) fractions after bevacizumab treatment (a). The levels of ET-1 mRNA (b) and ET-1 protein (c) were measured using real-time RT-PCR and the ELISA after bevacizumab and AS1842856 treatment. Data shown are means ± SD ( n = 3; ∗ p < 0.05). The experiments were repeated at least three times, with reproducible results.

Article Snippet: The concentration of ET-1 in the supernatant was determined by sandwich enzyme-linked immunosorbent assay (ELISA) using a human ET-1 Quantikine ELISA kit (R&D Systems, Abingdon, UK).

Techniques: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Quantitative RT-PCR

Journal: International Journal of Nephrology

Article Title: Bevacizumab Increases Endothelin-1 Production via Forkhead Box Protein O1 in Human Glomerular Microvascular Endothelial Cells In Vitro

doi: 10.1155/2021/8381115

Figure Lengend Snippet: Vascular endothelial growth factor A (VEGFA) is associated with the suppression of ET-1 production in cultured human glomerular microvascular endothelial cells (hGECs). hGECs were plated in 24-well plates (1 × 10 5 cells/well) for ELISA or 60 mm culture dishes (1 × 10 6 cells/dish) for RT-PCR. hGECs that were 90% confluent were serum-starved for 24 h before the experiments were performed. hGECs were treated with 0, 10, and 40 ng/mL of VEGFA or with a combination of 40 ng/mL VEGFA and 1 µ M bevacizumab for 8 h. The levels of ET-1 mRNA (a) and ET-1 protein (b) were measured using real-time RT-PCR and the ELISA after VEGFA and bevacizumab treatment. Data shown are means ± SD ( n = 3; ∗ p < 0.05). The experiments were repeated at least three times, with reproducible results.

Article Snippet: The concentration of ET-1 in the supernatant was determined by sandwich enzyme-linked immunosorbent assay (ELISA) using a human ET-1 Quantikine ELISA kit (R&D Systems, Abingdon, UK).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR