human brain endothelial cell line  (Cedarlane)

Journal: The European Journal of Neuroscience

Article Title: VEGF‐E Attenuates Injury After Ischemic Stroke by Promoting Reparative Revascularization

doi: 10.1111/ejn.70114

Figure Lengend Snippet: VEGF‐E increases basal PDGF‐D expression in brain endothelial cells by activating ERK1/2 pathway. (a) Schematic illustration of the experimental design of iHBEC stimulated with vehicle (VEH), VEGF‐E (100 ng/mL), or VEGF‐A (100 ng/mL) for 24 h, repeated for another 24 h under normoxic condition. (b) Representative Western blot images of PDGF‐D expression in iHBEC upon stimulation with VEH, VEGF‐E, or VEGF‐A. Analysis of the expression of (c) the full length (FL) and (d) the cleaved form of PDGF‐D. (e) Representative Western blot images of phosphorylated (p) and total (t) ERK1/2 expression in iHBEC upon stimulation with VEH, VEGF‐E, or VEGF‐A. Analysis of (f) p‐ERK1 and (g) t‐ERK1 expression as well as (h) p‐ERK1/t‐ERK1 ratio (i.e., pathway activation). Analysis of (i) p‐ERK2 and (j) t‐ERK2 expression as well as (k) p‐ERK2/t‐ERK2 ratio (i.e., pathway activation) in iHBEC upon stimulation with VEH, VEGF‐E, or VEGF‐A. Data are boxplot with min/max ( n = 3 independent experiments/condition). * p < 0.05/** p < 0.01 compared to control (c, f, h, one‐way ANOVA). Abbreviations: ANOVA, analysis of variance; iHBEC, immortalized human brain endothelial cells; PDGF, platelet‐derived growth factor; VEGF, vascular endothelial growth factor; VEH, vehicle.

Article Snippet: Immortalized human brain endothelial cells (iHBEC) (Cedarlane Laboratories, ON, Canada; CRL‐3245; 70041474) were used to investigate the molecular mechanisms underlying VEGF‐E action. iHBEC cells have all the characteristics needed to represent the BBB and interact with pericytes (Chen et al. ; Zolotoff et al. ).

Techniques: Expressing, Western Blot, Activation Assay, Control, Derivative Assay

Journal: The European Journal of Neuroscience

Article Title: VEGF‐E Attenuates Injury After Ischemic Stroke by Promoting Reparative Revascularization

doi: 10.1111/ejn.70114

Figure Lengend Snippet: VEGF‐E‐stimulated brain endothelial cells promote perivascular cell mobility under ischemia and reperfusion‐like condition. (a) Schematic illustration of the experimental design of iHBEC exposed to OGD and reperfusion followed by stimulation with vehicle (VEH) or VEGF‐E (100 ng/mL) for 24 h. (b) Representative Western blot images of phosphorylated (p) and total (t) ERK1/2 expression in iHBEC stimulated with vehicle (VEH) or VEGF‐E. Analysis of (c) p‐ERK1 and (d) t‐ERK1 expression as well as (e) p‐ERK1/t‐ERK1 ratio (i.e., pathway activation). Analysis of (f) p‐ERK2 and (g) t‐ERK2 expression as well as (h) p‐ERK2/t‐ERK2 ratio (i.e., pathway activation) in iHBEC stimulated with VEH and VEGF‐E after OGD. (i) Representative Western blot images of phosphorylated (p) and total (t) P38 expression. Analysis of (j) p‐P38 and (k) t‐P38 expression as well as (l) p‐P38/t‐P38 ratio (i.e., pathway activation) in iHBEC stimulated with VEH and VEGF‐E after OGD. (m) Schematic illustration of the experimental design of wound healing assay performed on HBVP after stimulation with VEH, VEGF‐E, or the conditioned medium (CM) of VEGF‐E‐treated iHBEC. (n) Representative brightfield images of the wound in VEH, VEGF‐E, or CM‐stimulated HBVP at baseline, 24 h, or 48 h. Analysis of wound closure of VEH, VEGF‐E, or CM‐treated HBVP at (o) 24 h and (p) 48 h, represented as percent of control (baseline, 0 h). Data are boxplot with min/max (n = 3–4 independent experiments/condition). * p < 0.05/** p < 0.01/*** p < 0.001 compared to control (c‐e, j, l, unpaired two‐tailed t ‐test; o, p, one‐way ANOVA). Abbreviations: H, hours; iHBEC, immortalized human brain endothelial cells; OGD, oxygen and glucose‐deprived; VEGF, vascular endothelial growth factor; VEH, vehicle.

Article Snippet: Immortalized human brain endothelial cells (iHBEC) (Cedarlane Laboratories, ON, Canada; CRL‐3245; 70041474) were used to investigate the molecular mechanisms underlying VEGF‐E action. iHBEC cells have all the characteristics needed to represent the BBB and interact with pericytes (Chen et al. ; Zolotoff et al. ).

Techniques: Western Blot, Expressing, Activation Assay, Wound Healing Assay, Control, Two Tailed Test