Journal: Journal of Animal Science and Technology

Article Title: Alterations in steroid hormone receptors, angiogenic, cell proliferation, apoptotic, and Ras-related factors during estrous cycle in porcine corpus luteum

doi: 10.5187/jast.2600012

Figure Lengend Snippet: List of antibodies for western blot

Article Snippet: Tie-2 , sc-293414 , Mouse , SCBT , 1:1000 , goat anti-mouse IgG-HRP , 1:10000.

Techniques: Western Blot

Journal: bioRxiv

Article Title: Ubiquitin ligase CHFR impairs Tie2 signaling via K 48 -linked ubiquitylation and degradation of Akt1 in endothelial cells

doi: 10.64898/2026.03.31.715582

Figure Lengend Snippet: a HMEC were transfected with scrambled-siRNA (sc-siRNA) or CHFR-siRNA. At 72 h post-transfection, cells stimulated with rAng1 (400 ng/ml) for different time intervals were used for IB analysis to determine phosphorylation of Akt1 at S473 and T308 ( n = 3 independent experiments). b Control HMEC and CHFR knockdown HMEC stimulated with rAng1 as above were stained with phospho-473-Akt1 specific antibody. c Control HMEC and CHFR knockdown HMEC were transfected with the Akt1 biosensor plasmid and then stimulated with rAng1 as above to assess live cell Akt1 activity by measuring the FRET ratio ( p <0.0001) ( left panel shows basal Akt activity; right panel shows Akt activity in response to rAng1 challenge). d-f Control HMEC and CHFR-siRNA treated HMEC stimulated with rAng1 were used for IB analysis to determine tyrosine phosphorylation Tie2 ( d ), phosphorylation of GSK3β at S9 and phosphorylation of β-catenin ( e ), ( n = 3 independent experiments). a, d-e Shown are mean values ± SEM ( n = 3 independent experiments; two-way ANOVA followed by Tukey’s post-hoc test).

Article Snippet: Rabbit polyclonal antibody (pAb) against phospho-Tie2 (catalog #AF3909; IB, 1:1000), goat polyclonal antibody (pAb) against PDGFRβ (catalog #AF1042; IS, 1:100) were from R&D system.

Techniques: Transfection, Phospho-proteomics, Control, Knockdown, Staining, Plasmid Preparation, Activity Assay

Journal: bioRxiv

Article Title: Ubiquitin ligase CHFR impairs Tie2 signaling via K 48 -linked ubiquitylation and degradation of Akt1 in endothelial cells

doi: 10.64898/2026.03.31.715582

Figure Lengend Snippet: a HMEC were transfected with WT or K/R-mutant Akt1 constructs. At 36 h after transfection, cells were treated with LPS (5 μg/ml) for 0, 12, and 24h and then cells were used for IB to determine expression of Akt1 and VE-cadherin. Shown are mean values ± SEM (n = 3 independent experiments; two-way ANOVA followed by Tukey’s post hoc test). b TIME endothelial cells (telomerase-immortalized human dermal microvascular endothelial cell line) were transfected with WT or K/R mutant Akt1 constructs and stimulated with LPS (5 μg/ml) for 0 and 6 h. Confocal imaging showed that expression of K/R mutant Akt1 prevents degradation of VE-cadherin. c WT mice were injected (i.v.) with liposome-encapsulated pmCherry-tagged WT or K/R mutant Akt1 constructs. Lungs harvested 96 h after injection were subjected to cryosection and stained with EC marker antibody vWF (green). Confocal imaging confirms expression of pmCherry-Akt1 (red) plasmid in lung endothelial cells. d-f Liposome-mediated delivery of Akt1 (WT) or K/R-mutated Akt1 in WT mice prevents degradation of VE-cadherin, mitigates LPS-induced lung vascular leak (EBA uptake), and reduces PMN transmigration (MPO assay). Shown are mean values ± SEM ( n = 3 or n = 5 mice/group; two-way ANOVA followed by Tukey’s post hoc test). g Model for E3 ligase CHFR regulation of endothelial junctional barrier integrity. Under baseline condition, constitutive Ang1-Tie2 signaling in EC maintains endothelial junctional barrier through Akt1 activation-mediated inhibition of the transcription factor FoxO1 activation and Ang-2 expression. During vascular inflammatory conditions such as sepsis, TLR4 signaling induces the expression of E3 ligase CHFR in a FoxO1-dependent manner. Then the upregulated CHFR mediates K 48 -linked polyubiquitylation and degradation of Akt1 and VE-cadherin (Tiruppathi et al., 2023) to disassemble EC junctional barrier. CHFR-mediated loss of FoxO1 negative regulator Akt1 expression in EC leads to increased FoxO1 expression which in turn promotes sustained expression of Ang-2 in EC to induce life-threatening pulmonary edema.

Article Snippet: Rabbit polyclonal antibody (pAb) against phospho-Tie2 (catalog #AF3909; IB, 1:1000), goat polyclonal antibody (pAb) against PDGFRβ (catalog #AF1042; IS, 1:100) were from R&D system.

Techniques: Transfection, Mutagenesis, Construct, Expressing, Imaging, Injection, Staining, Marker, Plasmid Preparation, Transmigration Assay, MPO Assay, Activation Assay, Inhibition

Journal: Inflammation and Regeneration

Article Title: Wnt10a exhibit spatiotemporal singularity in the temporal changes of angiogenesis in regenerated pulp-like tissue

doi: 10.1186/s41232-025-00397-y

Figure Lengend Snippet: Spatiotemporal specificity of angiogenesis and Tie-2 within regenerated dentin-like tissue after ectopic root transplantation

Article Snippet: Primary antibodies: Rabbit anti-Wnt10a (1:400) (NBP1-76,916, Novus Biologicals, CO, USA), anti-VEGF antibody (1:500) (19,003–1-AP, Proteintech, Chicago, IL, USA), anti-Tie-2 antibody (1:200) (19,157–1-AP, Proteintech, Chicago, IL, USA) were used.

Techniques: Transplantation Assay

Journal: Cell reports

Article Title: Angiopoietin-2 aggravates Alzheimer’s disease by promoting blood-brain barrier dysfunction and neuroinflammation

doi: 10.1016/j.celrep.2025.116621

Figure Lengend Snippet: (A and B) Reduced vascular lectin perfusion in the brain cortex of 6-month-old 5xFAD mice treated with ANGPT2 AAV compared to control-AAV-treated mice (A), with the corresponding quantification (B). Scale bars: 100 μm. White arrows indicate regions lacking a lectin signal in blood vessels. (C and D) Increased fibrinogen extravasation in the brain cortex of ANGPT2-AAV-treated 5xFAD mice compared to controls (C), with quantification (D). Arrowheads indicate extravasated fibrin. Scale bars: 50 μm. (E and F) Increased cadaverine deposition in the brain cortex of ANGPT2-AAV-treated 5xFAD mice compared to controls (E), with the corresponding quantification (F). Scale bars: 50 μm. (G–J) Decreased claudin-5 and occludin levels in ANGPT2-AAV-treated 5xFAD mice compared with controls (G and I), with quantification (H and J). Arrowheads indicate vessels negative for claudin-5 or occludin. Scale bars: 50 μm. (K–N) Increased PLVAP and caveolin-1 expression in cortical vessels of ANGPT2-AAV-treated 5xFAD mice compared with controls (K and M), with quantification (L and N). Scale bars: 50 μm. (O and P) Decreased pTIE2 immunoreactivity in ANGPT2-AAV-treated 5xFAD mice compared to controls (O), with the corresponding quantification (P). Arrowheads indicate reduced pTIE2 levels in blood vessels. Scale bars: 50 μm. Each dot represents an individual mouse in (B), (D), (F), (H), (J), (L), (N), and (P). Data are presented as the mean ± SEM.

Article Snippet: Next, sections were incubated overnight at 4°C with the following primary antibodies: For mouse tissue sections: CD31 (Thermo Fisher Scientific, MA3105), β-amyloid (Abcam, ab2539, ab126649), TIE2 (Regeneron Pharmaceuticals Inc., human monoclonal, REGN1376), pTIE2 (R&D systems, AF2720), ANGPT2 (Regeneron Pharmaceuticals Inc., human monoclonal, REGN910), PDGFR β (Thermo Fisher Scientific, 14-1402-82), fibrin(ogen) (Dako, A0080), claudin-5 (Thermo Fisher Scientific, 34-1600), GFAP (Aves Lab, GFAP), S100 β (Abcam, ab52642), Iba1 (Fujifilm Wako, 019-19741), CD74 (BioLegend, 151002), occludin (Thermo Fisher Scientific, 71-1500), PLVAP (BD Biosciences, 550563), and caveolin-1 (Santa Cruz Biotechnology, sc53564).

Techniques: Control, Expressing

Journal: Cell reports

Article Title: Angiopoietin-2 aggravates Alzheimer’s disease by promoting blood-brain barrier dysfunction and neuroinflammation

doi: 10.1016/j.celrep.2025.116621

Figure Lengend Snippet: (A) Experimental timeline for endothelial-specific ANGPT2 conditional knockout in 5xFAD mice (5xFAD;ANGPT2 iECKO ). Tamoxifen was administered at 2 months, and brains were harvested at 12 months of age. (B) No detectable ANGPT2 immunoreactivity in the blood vessels of the cortex in 12-month-old 5xFAD;ANGPT2 iECKO mice compared to 5xFAD;ANGPT2 WT mice. Scale bars: 100 μm. (C and D) Comparison of β-amyloid deposition in whole-brain cross-sections of 5xFAD;ANGPT2 iECKO and age-matched control 5xFAD mice (C) and its quantifications in the hippocampus and cortex (D). The yellow line outlines the brain boundary, and the yellow dotted line indicates the hippocampal regions. Scale bars: 1 mm. (E and F) Decreased fibrin leakage into the hippocampus and cortex of 5xFAD;ANGPT2 iECKO mice compared to controls (E), with corresponding quantification (F). Scale bars: 100 μm. (G and H) Enhanced endothelial pTIE2 levels in the hippocampus and cortex of 5xFAD;ANGPT2 iECKO mice compared to controls (G), with quantifications (H). Arrowheads indicate pTie2-expressing regions. Scale bars: 50 μm. (I and J) Decreased microglial activation in the hippocampus of 5xFAD;ANGPT2 iECKO mice compared to controls (I), with corresponding quantification in the hippocampus and cortex (J). Scale bars: 50 μm. Arrowheads indicate CD74 + activated microglia. Each dot represents an individual mouse in (D), (F), (H), and (J). Data are presented as the mean ± SEM.

Article Snippet: Next, sections were incubated overnight at 4°C with the following primary antibodies: For mouse tissue sections: CD31 (Thermo Fisher Scientific, MA3105), β-amyloid (Abcam, ab2539, ab126649), TIE2 (Regeneron Pharmaceuticals Inc., human monoclonal, REGN1376), pTIE2 (R&D systems, AF2720), ANGPT2 (Regeneron Pharmaceuticals Inc., human monoclonal, REGN910), PDGFR β (Thermo Fisher Scientific, 14-1402-82), fibrin(ogen) (Dako, A0080), claudin-5 (Thermo Fisher Scientific, 34-1600), GFAP (Aves Lab, GFAP), S100 β (Abcam, ab52642), Iba1 (Fujifilm Wako, 019-19741), CD74 (BioLegend, 151002), occludin (Thermo Fisher Scientific, 71-1500), PLVAP (BD Biosciences, 550563), and caveolin-1 (Santa Cruz Biotechnology, sc53564).

Techniques: Knock-Out, Comparison, Control, Expressing, Activation Assay