proteome profilertm human angiogenesis array kit  (R&D Systems)

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Endothelial PDGF-D contributes to neurovascular protection after ischemic stroke by rescuing pericyte functions

doi: 10.1007/s00018-024-05244-w

Figure Lengend Snippet: Brain pericyte survival and vascular protective properties are stimulated by PDGF-D. ( a ) Schematic illustration of the experimental design of primary human brain vascular pericytes (HBVP) exposition to oxygen and glucose deprivation (OGD) followed by rh-PDGF-D stimulation. ( b ) Western blot analysis showing BCL2 expression in vehicle (VEH)-, rh-PDGF-D 40 ng/mL (P40)- or rh-PDGF-D 80 ng/mL (P80)-treated HBVP, 24 h after OGD. ( c ) Western blot analysis showing NOTCH3 expression in VEH, P40 or P80 groups, 24 h after OGD. ( d ) Representative brightfield images of VEH and P80 groups, immediately upon or 24 h after wound formation (white dashed lines) under normoxic (NOR) or OGD conditions. ( e ) Analysis of wound closure of VEH and P80 groups under NOR or OGD conditions at 24 h, represented as percent of control (baseline, 0 h). ( f ) Representative images of the proteome Profiler human angiogenesis array membranes analyzing the expression of factors implicated in vascular remodeling in VEH and P80 groups exposed to NOR or OGD, 24 h after stimulation. Blue rectangles highlight downregulated factors and red rectangles highlight upregulated factors. ( g ) Heatmap illustration showing differentially regulated factors in OGD-exposed HBVP upon PDGF-D stimulation. ( h ) Analysis of VEGF, uPA and IGFBP1 expression in NOR- and OGD-exposed HBVP upon stimulation of PDGF-D. ( i ) Representative brightfield images of tubes formed by immortalized human brain endothelial cells (iHBEC) without treatment (EC), treated with P80 (EC + P80) or with CM of OGD-exposed HBVP (PC) stimulated with P80 (EC + CM), or co-cultured with HBVP (PC) without treatment (EC/PC), treated with P80 (EC/PC + P80) or with CM of OGD-exposed PC stimulated with P80 (EC/PC + CM), 8 h after plating cells on ECM. (j) Analysis of the density of tubes formed under the different conditions. ( b-e, j ) Data are boxplot with min/max ( n = 3–4/groups). * P < 0.05/ ** P < 0.01/ *** P < 0.001/ **** P < 0.0001 compared to other different condition ( (b) Kruskal-Wallis/Dunn’s test, ( c, e, j ) one-way ANOVA/Tukey’s test or ( h ) unpaired two tailed t-test). H, hour

Article Snippet: Proteome ProfilerTM human angiogenesis array kit (R&D Systems; ARY007) was used to simultaneously profile the expression of 55 angiogenesis-related proteins of unstimulated and PDGF-D-stimulated HBVP under normoxic or OGD conditions at 24 h, following manufacturer’s instructions.

Techniques: Western Blot, Expressing, Control, Cell Culture, Two Tailed Test

Journal: Viruses

Article Title: Human Brain Microvascular Endothelial Cells Exposure to SARS-CoV-2 Leads to Inflammatory Activation through NF-κB Non-Canonical Pathway and Mitochondrial Remodeling.

doi: 10.3390/v15030745

Figure Lengend Snippet: Figure 4. Production of angiogenic-related molecules is modulated by SARS-CoV-2 in HBMECs. (A) Conditioned medium from Mock and SARS-CoV-2-exposed HBMEC cultures (both with MOI 0.01 and 0.1) were analyzed via Proteome Profiler Human Angiogenic Antibody Array and detected by chemoluminescence, each protein detected in duplicated spots. (B) Densitometric analysis of membranes in (A) revealed the analytes with the strongest signal and which were affected by the SARS-CoV-2 challenge. Spots labelled 1-15 in (A) correspond to the analytes depicted in (B). (C) RT-qPCR analysis of angiogenesis-related genes in HBMECs revealed that PTX3 and HIF-1α were increased following SARS-CoV-2 exposure. *: p < 0.05; **: p < 0.01, two-way ANOVA with Bonferroni post-test of at least five independent experiments.

Article Snippet: Secretion of angiogenesis-related protein levels was detected using a Proteome ProfilerTM Human Angiogenesis Antibody Array kit (R&D Systems) according to the manufacturer’s instructions.

Techniques: Ab Array, Quantitative RT-PCR