brdu cell proliferation assay kit 200 test  (Advisains)

Journal: bioRxiv

Article Title: Identification of an osteopontin-derived peptide that binds neuropilin-1 and activates vascular cells

doi: 10.1101/2023.09.05.556237

Figure Lengend Snippet: The impact of FOL-026 on human umbilical vascular endothelial cells (HUVECs) proliferation (A) and apoptosis (B) induced with low-oxygen condition was determined by BrdU uptake (n=8-9 per group) and active caspase-3/-7 (n=7-10 per group) respectively. The reactive oxygen species (ROS) level (C) activated by 50ug/ml oxidized low-density lipoprotein (oxLDL) for 2 hours in cells pre-incubated with FOL-026 was assessed by H 2 O 2 measurement (n=4-8 per group). Representative pictures of endothelial migration into an in vitro injury model are shown (D) and the wound closure rate (E) in HUVECs stimulated with increasing concentration of FOL-026 was quantified by image J software (n=8-9 per group). The effect of FOL-026 on gene expression of vascular endothelial growth factor-A (VEGF-A, F ) and its receptors (G and H) , as well as the hepatocyte growth factor (HGF) receptor c-Met (I) in HUVECs (n=3-6 for F-I). Scale bar=150μm in D. Data are presented as means±SEM and acquired from 3 to 4 independent replicate tests. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 by one-way ANOVA and Dunnett’s post-hoc test.

Article Snippet: Cell proliferation was analyzed with BrdU Cell Proliferation Elisa Kit (Abcam, Cambridge, UK).

Techniques: Incubation, Migration, In Vitro, Concentration Assay, Software, Gene Expression

Journal: bioRxiv

Article Title: Identification of an osteopontin-derived peptide that binds neuropilin-1 and activates vascular cells

doi: 10.1101/2023.09.05.556237

Figure Lengend Snippet: (A) The efficiency of small interfering RNA transfection for 48 hours in HUVECs was confirmed by qRT-PCR (n=4). The effect of FOL-026 stimulation on cell proliferation (B) and cell apoptosis at 0.1% oxygen (C) in HUVECs transfected with siRNA-NC or siNRP-1 was evaluated by BrdU incorporation (n=4-7 per group) and active caspase-3/-7 (n=4-7 per group). Representative pictures of endothelial cells at 0 and 5 hours after induction of the wound are shown (D) . The wound closure rate (E) of siRNA-NC/ siNRP-1-transfected HUVECs stimulated with increasing concentration of FOL-026 was quantified by image J software (n=9-12 per group). Scale bar =150μm in D. Data are presented as means ± SEM and acquired from 3 to 4 independent replicate tests. Two-way ANOVA followed with Sidak’s multiple comparisons test for A, two-way ANOVA followed with Sidak’s and Dunnett’s multiple comparisons test for B, C and E.*P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001.

Article Snippet: Cell proliferation was analyzed with BrdU Cell Proliferation Elisa Kit (Abcam, Cambridge, UK).

Techniques: Small Interfering RNA, Transfection, Quantitative RT-PCR, BrdU Incorporation Assay, Concentration Assay, Software

Journal: bioRxiv

Article Title: Identification of an osteopontin-derived peptide that binds neuropilin-1 and activates vascular cells

doi: 10.1101/2023.09.05.556237

Figure Lengend Snippet: The effect of FOL-026 treatment on smooth muscle cell proliferation (A) and apoptosis (B) induced by low-oxygen condition were evaluated with the uptake of BrdU (n=10 per group) and active caspase-3/-7 (n=6-8 per group) respectively. H 2 O 2 measurement (C) in smooth muscle cells pre-incubated with FOL-26 peptides following 50μg/ml oxLDL exposure for 2 hours (n=6-7 per group). Representative images of the in vitro wound assay are shown (D) and the ability of wound closure (E) in human coronary smooth muscle cells (HCASMCs) treated with different concentrations of FOL-026 was measured by image J software (n=10 per group). qRT-PCR analysis of collagen type I α 1 chain (COL1A1, F ) and collagen type I α 2 chain (COL1A2, G ) mRNA expression in FOL-026 peptide-stimulated HCASMCs (n=6 for F and G). Western blot analysis of phosphorylated AKT (p-AKT-T308 and p-AKT-S473), t-AKT, phosphorylated ERK1/2 (p-ERK1/2) and t-ERK1/2 in HCASMCs stimulated with FOL-026 (H-J) . Quantification of Akt and ERK1/2 phosphorylation were normalized to respective total protein expressions (n=7 for H, n=9 for I, n=3 for J). Scale bar =150μm in D. Data are presented as means ± SEM and acquired from 3 to 4 independent replicate tests. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 by one-way ANOVA and Dunnett’s post-hoc test.

Article Snippet: Cell proliferation was analyzed with BrdU Cell Proliferation Elisa Kit (Abcam, Cambridge, UK).

Techniques: Incubation, In Vitro, Software, Quantitative RT-PCR, Expressing, Western Blot, Phospho-proteomics