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anti vegfr2 oti12c1  (OriGene)
94
OriGene anti vegfr2 oti12c1
(A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) <t>with</t> <t>anti-VEGFR2</t> antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.
Anti Vegfr2 Oti12c1, supplied by OriGene, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+vegf+receptor/VEGF+Receptor+2+(KDR)+Mouse+Monoclonal+Antibody/bio_rxiv__64898__2026__05__12__724648-193-18-21
Average 94 stars, based on 1 article reviews
anti vegfr2 oti12c1 - by Bioz Stars, 2026-09
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rabbit anti p irf3  (Cell Signaling Technology Inc)
93
Cell Signaling Technology Inc rabbit anti p irf3
(A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) <t>with</t> <t>anti-VEGFR2</t> antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.
Rabbit Anti P Irf3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+vegf+receptor/VEGF+Receptor+2+Control+Proteins/pmc13035991-56-32-36
Average 93 stars, based on 1 article reviews
rabbit anti p irf3 - by Bioz Stars, 2026-09
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239 anti vegfr2 py1175  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc 239 anti vegfr2 py1175
(A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) <t>with</t> <t>anti-VEGFR2</t> antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.
239 Anti Vegfr2 Py1175, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+vegf+receptor/Phospho-VEGF+Receptor+2+(Tyr1175)+Rabbit+mAb/10__1158_slash_2767___9764__crc___25___0634-135-19-23
Average 95 stars, based on 1 article reviews
239 anti vegfr2 py1175 - by Bioz Stars, 2026-09
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anti 238 vegfr2  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti 238 vegfr2
(A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) <t>with</t> <t>anti-VEGFR2</t> antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.
Anti 238 Vegfr2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+vegf+receptor/VEGF+Receptor+2+Rabbit+mAb/10__1158_slash_2767___9764__crc___25___0634-135-8-11
Average 96 stars, based on 1 article reviews
anti 238 vegfr2 - by Bioz Stars, 2026-09
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anti vegf receptor 2  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti vegf receptor 2
(A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) <t>with</t> <t>anti-VEGFR2</t> antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.
Anti Vegf Receptor 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+vegf+receptor/VEGF+Receptor+2+Rabbit+mAb/10__1113_slash_jp290159-123-30-35
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anti vegf receptor 2 - by Bioz Stars, 2026-09
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anti vegfr2  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc anti vegfr2
(A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) <t>with</t> <t>anti-VEGFR2</t> antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.
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vegfr2 rabbit mab  (Cell Signaling Technology Inc)
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Re suppresses ox-LDL-induced HUVEC proliferation and migration. (A, B) Results of the CCK-8 assay, n = 6. (C) Representative images of HUVECs at 0 h and 12 h after ox-LDL induction in wound healing experiments, bar = 50 μm. (D) Quantification of EC migration in the wound healing assay, n = 3. (E) Western blot assay and quantitative data of VE-cadherin and <t>VEGFR2</t> in HUVECs, n = 3. ### p < 0.001, ## p < 0.01, # p < 0.05, vs. control group, ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05, vs. ox-LDL group.
Vegfr2 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibody vegfr2  (Cell Signaling Technology Inc)
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(A) HUVECs were infected with Ad-Moesin WT or Ad-Moesin T558D for 48 hrs, and total RNA was harvested for RNA-seq. The differentially expressed genes were analyzed with Gene Ontology Enrichment Analysis. The top nine gene sets are shown. Data were analyzed from GSE302986. (B) HUVECs transfected with siCtrl or si KINDLIN-2 were serum-starved (2% FBS) for 8 hrs, then stimulated with 2 μg/mL Alexa 594-labeled VEGF (red) for 30 mins. VEGF accumulation was visualized by confocal microscopy. Actin, phalloidin (green); nuclei, DAPI (blue). (C) Quantification of Alexa 594-labeled VEGF spots normalized to cellular area (n= 4 independent experiments). (D) HUVECs infected with Ad-Moesin WT or Ad-Moesin T558D were starved and stimulated with Alexa 594-labeled VEGF as in (B). VEGF accumulation was imaged by confocal microscopy. (E) Quantification of Alexa 594-labeled VEGF spots normalized to cellular area (n=3 independent experiments). (F) HUVECs transfected with siCtrl or si KINDLIN-2 were serum-starved for 8 hrs, stimulated with 50 ng/ml VEGF for 30 mins, fixed, and stained for <t>VEGFR2</t> (green). Representative images show intracellular VEGFR2 vesicles. (G) Quantification of VEGFR2 vesicle number per cell from (F) (n=3 independent experiments). (H) HUVECs infected with Ad-Moesin WT or Ad-Moesin T558D were treated as in (F), and stained for VEGFR2. (I) Quantification of VEGFR2 vesicle number per cell from (H) (n= 3 independent experiments). (J) Schematic of the cell surface biotinylation assay. HUVECs were starved for 8 hrs, labeled with EZ-Link Sulfo-NHS-SS-Biotin (0.25 mg/mL) at 4 °C for 1 hr, then stimulated with VEGF (50 ng/mL) for 30 mins. After surface biotin was stripped with GSH elution buffer, total proteins were extracted. Biotinylated internalized VEGFR2 was pulled down using streptavidin magnetic beads and analyzed by western blotting. (K) Cell surface biotinylation assay for VEGFR2 internalization in siCtrl- and si KINDLIN-2 –transfected HUVECs. Input lysates show VEGFR2, Kindlin-2, and GAPDH. “Surf” represents surface VEGFR2 prior to VEGF stimulation and biotin stripping. (L) Quantification of internalized VEGFR2 normalized to surface VEGFR2 levels (n=4 independent experiments). (M) Schematic of in vivo Alexa 594-labeled VEGF uptake assay in Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. Tamoxifen was administered from P1 to P3; Alexa 594-labeled VEGF was injected intravitreally at P6 and analyzed after 30 mins. (N) Representative confocal images of the retinal angiogenic front showing uptake of Alexa 594-labeled VEGF by ECs from Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. (O) Quantification of internalized Alexa 594-VEGF at the angiogenic front (n = 4/4 pups). (P) Western blot of HUVECs transfected with siCtrl or si KINDLIN-2 , starved for 8 hrs, then stimulated with 50 ng/mL VEGF. (Q) Quantification of p-ERK levels at 5 mins after VEGF stimulation from (P) (n = 4 independent experiments). (R) Western blot of HUVECs infected with Ad-Vector, Ad-Moesin WT , or Ad-Moesin T558D , treated as in (P). (S) Quantification of p-ERK levels at 5 mins after VEGF stimulation from (R) (n = 3 independent experiments). Data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 by two-tailed Student’s t-test or one-way ANOVA followed by Tukey’s multiple comparisons test. Scale bars: 5 μm in (B), (D), (F), (H), and (N).
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(A) Volcano plot of differentially secreted proteins from primary LSECs treated with 40 ng/mL LIF versus control (n = 4). (B&C) qPCR of lifr (B) and hgf (C) levels in LSECs isolated from lifr f/f mice with indicated treatments. (D) HGF concentrations in conditioned medium from LSECs treated with hLIF at the indicated doses. (E) HGF concentrations in conditioned medium from LSECs treated with 40 ng/mL hLIF in the presence of the <t>STAT3</t> inhibitor C188-9 at the indicated doses. (F) HGF concentrations in conditioned medium from LSECs isolated from lifr f/f or lifrΔEC mice treated with hLIF, hCT-1 or hOSM. (G) Western blot analysis of hepatocytes co-cultured with LSECs with or without 40 ng/mL LIF. Quantification is shown in Figure S4C. (H) Model depicting LIF action on LSECs to stimulate angiogenesis and HGF release, thereby supporting hepatocyte proliferation via paracrine regulation.
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(A) Volcano plot of differentially secreted proteins from primary LSECs treated with 40 ng/mL LIF versus control (n = 4). (B&C) qPCR of lifr (B) and hgf (C) levels in LSECs isolated from lifr f/f mice with indicated treatments. (D) HGF concentrations in conditioned medium from LSECs treated with hLIF at the indicated doses. (E) HGF concentrations in conditioned medium from LSECs treated with 40 ng/mL hLIF in the presence of the <t>STAT3</t> inhibitor C188-9 at the indicated doses. (F) HGF concentrations in conditioned medium from LSECs isolated from lifr f/f or lifrΔEC mice treated with hLIF, hCT-1 or hOSM. (G) Western blot analysis of hepatocytes co-cultured with LSECs with or without 40 ng/mL LIF. Quantification is shown in Figure S4C. (H) Model depicting LIF action on LSECs to stimulate angiogenesis and HGF release, thereby supporting hepatocyte proliferation via paracrine regulation.
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Image Search Results


(A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) with anti-VEGFR2 antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) with anti-VEGFR2 antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Mutagenesis, Generated, Software, Injection, Isolation, Immunoprecipitation, Control, Western Blot, Marker, Staining, Fluorescence

(A), HEK293T cells transfected with either pCMV3 vector or VEGFR2-Myc-expressing vector were treated with either vehicle or RO overnight as in 1F. Membrane fractions were prepared as in Methods and incubated with vehicle (-) or VEGF-A as in 1F for 30 minutes in the presence of lactacystin. VEGFR2-Myc, VCTF1-Myc, and VCTF2-Myc, were then detected on WB using anti-Myc antibodies. Representative blot shows Myc-labelled fragments as indicated in Figure. (B), Left: HEK293T cells were transfected with VEGFR2-Myc and treated with vehicle or RO as in 3A and extracted in SDS buffer as in methods. VEGFR2-Myc and VCTF1-Myc were detected in cell extracts on WBs with antibodies recognizing the cytoplasmic sequence of VEGFR2 (ab39256). β-actin: loading control. Right: HEK293T cells were transfected as in 3A. Cells were pretreated with 200nm ADAM17 inhibitor D1 (A12: ADAM17 inh) for 1 hour and then stimulated with vehicle or VEGF-A as in 3A for 1h. Cells were extracted as in 3B Left. VEGFR2-Myc and its proteolytic product VCTF1-Myc are detected with anti-Myc antibody on WB as in 3A. Vinculin: loading control. (C), Left: WT pCECs were treated with either vehicle (DMSO; Veh) or RO as in 1F for 15-16 h and extracted in SDS buffer. VEGFR2 and VCTF1 were detected in cell extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. Right: Brain MVs were isolated from adult WT mice as in 1B following 15 hours of treatment with either vehicle or RO as in 1D and extracted in SDS buffer as in Methods. VEGFR2 and VCTF1 were detected in MV extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. (D), Left: pCECs isolated from WT or PS1 knockout (PS1 KO) mouse embryos were extracted as in 3C. VEGFR2 and VCTF1 were detected in cell extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. Right: HEK293T cells expressing VEGFR2-Myc were transfected with anti-PS1 siRNA or non-targeting control siRNA (NT siRNA) as in Methods and extracted as in 3B. VEGFR2-Myc and VCTF1-Myc were detected in cell extracts with anti-Myc antibody as in 3A. PS1 N-terminal fragment (PS1/NTF) was detected in cell extracts with anti-PS1 antibody (R222; 41). β-actin: loading control. (E), Left: Extracts from WT and either WT/M146V- or WT/I213T-expressing pCECs were prepared as in 3C. VEGFR2 and VCTF1 were detected in cell extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. Right: Brain MVs were isolated from WT and either WT/M146V- or WT/I213T-expressing mice as in 1B and extracted as in 3C Right. VEGFR2 and VCTF1 were detected in MV extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. Each WB is representative of at least three independent experiments.

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A), HEK293T cells transfected with either pCMV3 vector or VEGFR2-Myc-expressing vector were treated with either vehicle or RO overnight as in 1F. Membrane fractions were prepared as in Methods and incubated with vehicle (-) or VEGF-A as in 1F for 30 minutes in the presence of lactacystin. VEGFR2-Myc, VCTF1-Myc, and VCTF2-Myc, were then detected on WB using anti-Myc antibodies. Representative blot shows Myc-labelled fragments as indicated in Figure. (B), Left: HEK293T cells were transfected with VEGFR2-Myc and treated with vehicle or RO as in 3A and extracted in SDS buffer as in methods. VEGFR2-Myc and VCTF1-Myc were detected in cell extracts on WBs with antibodies recognizing the cytoplasmic sequence of VEGFR2 (ab39256). β-actin: loading control. Right: HEK293T cells were transfected as in 3A. Cells were pretreated with 200nm ADAM17 inhibitor D1 (A12: ADAM17 inh) for 1 hour and then stimulated with vehicle or VEGF-A as in 3A for 1h. Cells were extracted as in 3B Left. VEGFR2-Myc and its proteolytic product VCTF1-Myc are detected with anti-Myc antibody on WB as in 3A. Vinculin: loading control. (C), Left: WT pCECs were treated with either vehicle (DMSO; Veh) or RO as in 1F for 15-16 h and extracted in SDS buffer. VEGFR2 and VCTF1 were detected in cell extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. Right: Brain MVs were isolated from adult WT mice as in 1B following 15 hours of treatment with either vehicle or RO as in 1D and extracted in SDS buffer as in Methods. VEGFR2 and VCTF1 were detected in MV extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. (D), Left: pCECs isolated from WT or PS1 knockout (PS1 KO) mouse embryos were extracted as in 3C. VEGFR2 and VCTF1 were detected in cell extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. Right: HEK293T cells expressing VEGFR2-Myc were transfected with anti-PS1 siRNA or non-targeting control siRNA (NT siRNA) as in Methods and extracted as in 3B. VEGFR2-Myc and VCTF1-Myc were detected in cell extracts with anti-Myc antibody as in 3A. PS1 N-terminal fragment (PS1/NTF) was detected in cell extracts with anti-PS1 antibody (R222; 41). β-actin: loading control. (E), Left: Extracts from WT and either WT/M146V- or WT/I213T-expressing pCECs were prepared as in 3C. VEGFR2 and VCTF1 were detected in cell extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. Right: Brain MVs were isolated from WT and either WT/M146V- or WT/I213T-expressing mice as in 1B and extracted as in 3C Right. VEGFR2 and VCTF1 were detected in MV extracts on WB with anti-VEGFR2 antibody as in 3B Left. β-actin: loading control. Each WB is representative of at least three independent experiments.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Transfection, Plasmid Preparation, Expressing, Membrane, Incubation, Sequencing, Control, Isolation, Knock-Out

(A), HEK293T cells were transfected with VEGFR2-Myc as in 3A and treated with vehicle or VEGF-A for the indicated times, in the presence or absence of RO as in 1F and extracted in SDS buffer. Left: VEGFR2 dimer and monomer were detected on WB with anti-Myc antibodies. β-actin: loading control. Right: Graph shows the fold change in the VEGFR2 dimer to monomer ratio. (B), WT mice were treated with vehicle or RO as in 1D. Mice were treated with VEGF-A, and brain MVs were isolated and extracted as in 1B. Left: VEGFR2 dimers and monomers were detected on WB with anti-VEGFR2 antibody D5B1. Vinculin: loading control. Right: Graph shows the fold change in the VEGFR2 dimer to monomer ratio. (C), WT and WT/M146V or WT/I213T mice were injected with either vehicle or VEGF-A via the carotid artery as in 1D, and brain MVs were prepared as in 1B and extracted as in 3C Right. Left: VEGFR2 dimers and monomers were detected on WB with anti-VEGFR2 antibody as in 4B Left. β-actin: loading control. Right: graph shows the fold change in the VEGFR2 dimer to monomer ratio. (D), HEK293T cells were co-transfected with VEGFR2-Myc as in 3A and either WT PS1 or PS1 mutant M146V or I213T in FCbAIGW vector as indicated in Figure. Cells were treated with VEGF-A as in 1G for the indicated times and extracted in SDS buffer. Left: VEGFR2 dimers and monomers were detected in cell extract on WB with anti-Myc antibody as in 4A. Full length PS1 (FL-PS1) and PS1/NTF were detected with R222 (middle). Vinculin: loading control. Right: Graph shows the fold change in the VEGFR2 dimer to monomer ratio. (E), HEK293 cells were co-transfected with VEGFR2-Myc and either non-targeting or anti-PS1 siRNA as in 3D. Cells were treated with vehicle (0 lanes) or VEGF-A as in 4D above for the indicated times and extracted in SDS buffer. Left: VEGFR2 dimers and monomers and VCTF1 were detected on WB with anti-Myc antibody as in 4A. PS1/NTF was detected in cell extracts with R222. β-actin: loading control. Right: graphs show fold change in PS1/NTF levels (upper) and fold change in the VEGFR2 dimer/monomer ratio (lower) following treatment with anti-PS1 siRNA. PS1 downregulation resulted in decreased VEGFR2 dimerization and increased VCTF1-Myc (upper panel). (F), Embryonic brain (E15.5) extract from WT or PS1 knockout (PS1 KO) mice were prepared as described . Representative WB of extracts shows VEGFR2 dimers, detected with anti-VEGFR2 antibody D5B1, and PS1-NTF detected with R222 antibody. Vinculin: loading control. A–E: data are presented as mean ± SE from at least three independent experiments. Statistical analysis was performed by two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A), HEK293T cells were transfected with VEGFR2-Myc as in 3A and treated with vehicle or VEGF-A for the indicated times, in the presence or absence of RO as in 1F and extracted in SDS buffer. Left: VEGFR2 dimer and monomer were detected on WB with anti-Myc antibodies. β-actin: loading control. Right: Graph shows the fold change in the VEGFR2 dimer to monomer ratio. (B), WT mice were treated with vehicle or RO as in 1D. Mice were treated with VEGF-A, and brain MVs were isolated and extracted as in 1B. Left: VEGFR2 dimers and monomers were detected on WB with anti-VEGFR2 antibody D5B1. Vinculin: loading control. Right: Graph shows the fold change in the VEGFR2 dimer to monomer ratio. (C), WT and WT/M146V or WT/I213T mice were injected with either vehicle or VEGF-A via the carotid artery as in 1D, and brain MVs were prepared as in 1B and extracted as in 3C Right. Left: VEGFR2 dimers and monomers were detected on WB with anti-VEGFR2 antibody as in 4B Left. β-actin: loading control. Right: graph shows the fold change in the VEGFR2 dimer to monomer ratio. (D), HEK293T cells were co-transfected with VEGFR2-Myc as in 3A and either WT PS1 or PS1 mutant M146V or I213T in FCbAIGW vector as indicated in Figure. Cells were treated with VEGF-A as in 1G for the indicated times and extracted in SDS buffer. Left: VEGFR2 dimers and monomers were detected in cell extract on WB with anti-Myc antibody as in 4A. Full length PS1 (FL-PS1) and PS1/NTF were detected with R222 (middle). Vinculin: loading control. Right: Graph shows the fold change in the VEGFR2 dimer to monomer ratio. (E), HEK293 cells were co-transfected with VEGFR2-Myc and either non-targeting or anti-PS1 siRNA as in 3D. Cells were treated with vehicle (0 lanes) or VEGF-A as in 4D above for the indicated times and extracted in SDS buffer. Left: VEGFR2 dimers and monomers and VCTF1 were detected on WB with anti-Myc antibody as in 4A. PS1/NTF was detected in cell extracts with R222. β-actin: loading control. Right: graphs show fold change in PS1/NTF levels (upper) and fold change in the VEGFR2 dimer/monomer ratio (lower) following treatment with anti-PS1 siRNA. PS1 downregulation resulted in decreased VEGFR2 dimerization and increased VCTF1-Myc (upper panel). (F), Embryonic brain (E15.5) extract from WT or PS1 knockout (PS1 KO) mice were prepared as described . Representative WB of extracts shows VEGFR2 dimers, detected with anti-VEGFR2 antibody D5B1, and PS1-NTF detected with R222 antibody. Vinculin: loading control. A–E: data are presented as mean ± SE from at least three independent experiments. Statistical analysis was performed by two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Transfection, Control, Isolation, Injection, Mutagenesis, Plasmid Preparation, Knock-Out

(A), HEK293T cells were co-transfected with both constructs expressing complementary VEGFR2-Venus fusion proteins as in Methods. Cells were treated with vehicle or VEGF-A for 30 minutes in the presence or absence of RO as in 1F and imaged as in Methods. Left upper panel: Green fluorescence indicates VEGFR2 homodimer formation. Scale bar: 200μm. Left lower panel: Corresponding phase contrast images of cell cultures. Right: Graph shows fold induction of VEGFR2 homodimer formation based on fluorescent intensity measured using ImageJ as in Methods. (B), HEK293T cells were co-transfected with VEGFR2-Myc in pCMV3 vector and either WT PS1 or one of the PS1 FAD mutants indicated in Figure in FCbAIGW. Cells were treated with vehicle or VEGF-A for 0, 15 or 30 mins and then extracted in SDS buffer as in Methods. Left: VEGFR2 dimers and monomers were detected on WB with anti-VEGFR2 antibody (OTI12C1). PS1 (FL-PS1) and PS1/NTF were detected with R222. Vinculin: loading control. Right: Graph shows fold change in the VEGFR2 dimer to monomer ratio. For statistical analysis, two-way ANOVA followed by Tukey post-hoc test was used. ns = not significant, **p<0.01, ***p<0.001.

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A), HEK293T cells were co-transfected with both constructs expressing complementary VEGFR2-Venus fusion proteins as in Methods. Cells were treated with vehicle or VEGF-A for 30 minutes in the presence or absence of RO as in 1F and imaged as in Methods. Left upper panel: Green fluorescence indicates VEGFR2 homodimer formation. Scale bar: 200μm. Left lower panel: Corresponding phase contrast images of cell cultures. Right: Graph shows fold induction of VEGFR2 homodimer formation based on fluorescent intensity measured using ImageJ as in Methods. (B), HEK293T cells were co-transfected with VEGFR2-Myc in pCMV3 vector and either WT PS1 or one of the PS1 FAD mutants indicated in Figure in FCbAIGW. Cells were treated with vehicle or VEGF-A for 0, 15 or 30 mins and then extracted in SDS buffer as in Methods. Left: VEGFR2 dimers and monomers were detected on WB with anti-VEGFR2 antibody (OTI12C1). PS1 (FL-PS1) and PS1/NTF were detected with R222. Vinculin: loading control. Right: Graph shows fold change in the VEGFR2 dimer to monomer ratio. For statistical analysis, two-way ANOVA followed by Tukey post-hoc test was used. ns = not significant, **p<0.01, ***p<0.001.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Transfection, Construct, Expressing, Fluorescence, Plasmid Preparation, Control

(A), pCECs from WT or HTRZ (WT/M146V or WT/I213T) or HMZG (M146V/M146V or I213T/I213T) mice were extracted in SDS buffer as in Methods. VEGFR2 and VCTF1 were detected on WBs of cell extracts as in 3B with the anti-VEGFR2 antibody (ab39256). β-actin: loading control. (B), Brain MVs were isolated from adult WT mice or mice HTRZ or HMZG for PS1 FAD mutants M146V or I213T (see Suppl. 5A). VEGFR2 and VCTF1 were detected in MV extracts on WB as in Suppl. 5A with anti-VEGFR2 antibody (ab39256). β-actin: loading control.

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A), pCECs from WT or HTRZ (WT/M146V or WT/I213T) or HMZG (M146V/M146V or I213T/I213T) mice were extracted in SDS buffer as in Methods. VEGFR2 and VCTF1 were detected on WBs of cell extracts as in 3B with the anti-VEGFR2 antibody (ab39256). β-actin: loading control. (B), Brain MVs were isolated from adult WT mice or mice HTRZ or HMZG for PS1 FAD mutants M146V or I213T (see Suppl. 5A). VEGFR2 and VCTF1 were detected in MV extracts on WB as in Suppl. 5A with anti-VEGFR2 antibody (ab39256). β-actin: loading control.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Control, Isolation

(A), HEK293T cells were co-transfected with vector expressing VEGFR2-Myc and plasmids expressing EphB2/CTF1-Flag, VCTF1-Flag, or vector alone. Cells were stimulated with vehicle (-) or VEGF-A (+) as in 1F and then extracted in SDS buffer. Upper panel: VEGFR2 monomers and dimers are detected on WBs using anti-VEGFR2 antibody OTI12C1. Middle panel: VCTF1-Flag and EphB2/CTF1-Flag expression was detected using anti-Flag antibody. Lower panel: GAPDH, loading control. Graph shows fold change of the VEGFR2 dimer to monomer ratio. (B), HEK293T cells were co-transfected with vector expressing VEGFR2-Myc and either vector alone or increasing amounts of vector expressing VCTF1-Flag (200ng, 400ng, 800ng, or 1000ng). Forty-eight hours post-transfection cells were stimulated with either vehicle or VEGF-A as in 1F for 20 min and extracted in SDS buffer. Upper panel: VEGFR2 dimers and monomers (VEGFR2-Myc) are detected in cell lysates on WBs using anti-VEGFR2 antibody OTI12C1. Middle panel: VCTF1-Flag is detected with anti-Flag antibody. Lower panel: GAPDH: loading control. Bottom: Non-linear regression analysis (inhibitor vs. response, three-parameter model) showed a good fit to the curve (R² = 0.7410), indicating that increase of VCTF1-Flag expression inhibits VEGF-A-induced VEGFR2 dimerization. The red line (squares) represents VEGF-A-treated cells, whereas the blue line (circles) represents vehicle-treated cells. (C), Cells described in 5A were lysed in Triton X-100 buffer as in Methods and lysates were IPed with anti-Flag antibody. Upper panel: VEGFR2-Myc co-IPed with VCTF1-Flag is detected on WB using anti-Myc antibody as in 3A. Second panel: IPed VCTF1-Flag and EphB2/CTF1-Flag are detected on WBs with anti-Flag antibody. Third panel: Input of VEGFR2-Myc is detected with anti-Myc antibody as in 3A. Fourth panel: Input VCTF1-Flag and EphB2/CTF1-Flag are detected with Flag antibody. GAPDH: loading control. (D), bEnd3 cells were transduced with lentiviral vector FCbAIGW expressing VCTF1-Flag or empty vector as in Methods. Cells were treated with vehicle (-) or VEGF-A (+) as in Suppl. 4A for 7 minutes and extracted in SDS buffer as in Methods. Left: VEGFR2 dimers, monomers, p-VEGFR2 (Tyr1175), p-ERK1/2, ERK1/2 and VCTF1-Flag are detected on WBs with specific antibodies. Right: Graphs show fold change of VEGFR2 dimer to monomer ratio or p-VEGFR2/VEGFR2 and p-ERK1/2/ERK1 protein ratios. (E), bend3 cells expressing either empty vector (FCbAIGW) or VCTF1-Flag as in 5D were seeded as in Methods and treated with vehicle (-) or VEGF-A (+) as in Suppl. 4A for 6 hours. Upper: Representative photomicrographs show tube-like (loop/mesh) structures. Fluorescent images (EGFP, green) are shown. Scale bar 200μm. Lower: Graph shows quantification of tube formation as average number of loops/meshes per field. For statistical analysis, two-way ANOVA followed by Tukey post-hoc test was performed. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A), HEK293T cells were co-transfected with vector expressing VEGFR2-Myc and plasmids expressing EphB2/CTF1-Flag, VCTF1-Flag, or vector alone. Cells were stimulated with vehicle (-) or VEGF-A (+) as in 1F and then extracted in SDS buffer. Upper panel: VEGFR2 monomers and dimers are detected on WBs using anti-VEGFR2 antibody OTI12C1. Middle panel: VCTF1-Flag and EphB2/CTF1-Flag expression was detected using anti-Flag antibody. Lower panel: GAPDH, loading control. Graph shows fold change of the VEGFR2 dimer to monomer ratio. (B), HEK293T cells were co-transfected with vector expressing VEGFR2-Myc and either vector alone or increasing amounts of vector expressing VCTF1-Flag (200ng, 400ng, 800ng, or 1000ng). Forty-eight hours post-transfection cells were stimulated with either vehicle or VEGF-A as in 1F for 20 min and extracted in SDS buffer. Upper panel: VEGFR2 dimers and monomers (VEGFR2-Myc) are detected in cell lysates on WBs using anti-VEGFR2 antibody OTI12C1. Middle panel: VCTF1-Flag is detected with anti-Flag antibody. Lower panel: GAPDH: loading control. Bottom: Non-linear regression analysis (inhibitor vs. response, three-parameter model) showed a good fit to the curve (R² = 0.7410), indicating that increase of VCTF1-Flag expression inhibits VEGF-A-induced VEGFR2 dimerization. The red line (squares) represents VEGF-A-treated cells, whereas the blue line (circles) represents vehicle-treated cells. (C), Cells described in 5A were lysed in Triton X-100 buffer as in Methods and lysates were IPed with anti-Flag antibody. Upper panel: VEGFR2-Myc co-IPed with VCTF1-Flag is detected on WB using anti-Myc antibody as in 3A. Second panel: IPed VCTF1-Flag and EphB2/CTF1-Flag are detected on WBs with anti-Flag antibody. Third panel: Input of VEGFR2-Myc is detected with anti-Myc antibody as in 3A. Fourth panel: Input VCTF1-Flag and EphB2/CTF1-Flag are detected with Flag antibody. GAPDH: loading control. (D), bEnd3 cells were transduced with lentiviral vector FCbAIGW expressing VCTF1-Flag or empty vector as in Methods. Cells were treated with vehicle (-) or VEGF-A (+) as in Suppl. 4A for 7 minutes and extracted in SDS buffer as in Methods. Left: VEGFR2 dimers, monomers, p-VEGFR2 (Tyr1175), p-ERK1/2, ERK1/2 and VCTF1-Flag are detected on WBs with specific antibodies. Right: Graphs show fold change of VEGFR2 dimer to monomer ratio or p-VEGFR2/VEGFR2 and p-ERK1/2/ERK1 protein ratios. (E), bend3 cells expressing either empty vector (FCbAIGW) or VCTF1-Flag as in 5D were seeded as in Methods and treated with vehicle (-) or VEGF-A (+) as in Suppl. 4A for 6 hours. Upper: Representative photomicrographs show tube-like (loop/mesh) structures. Fluorescent images (EGFP, green) are shown. Scale bar 200μm. Lower: Graph shows quantification of tube formation as average number of loops/meshes per field. For statistical analysis, two-way ANOVA followed by Tukey post-hoc test was performed. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Transfection, Plasmid Preparation, Expressing, Control, Transduction

(A), Brain tissue extracts were prepared as in Methods from twelve PS1 FAD patients each carrying a different PS1 mutation, and twelve non-demented controls described in Methods. Left: VEGFR2 dimers and monomers were detected in brain extracts on WBs using anti-VEGFR2 antibody D5B1. Representative gels with control (C1-4) or FAD samples (FAD1-4) expressing mutants P264L, A260V, N135S and P242H respectively are shown. Vinculin: loading control. Right: Graph shows the fold change in VEGFR2 dimer to monomer ratio of FAD and control samples. (B), Brain tissue extract from control and PS1 FAD patient brains described in 7A were prepared and IPed with anti-endoglin antibody (ab252345) or IgG as in Methods. Upper panel: VEGFR2 co-IPed with endoglin was detected on WBs using an anti-VEGFR2 antibody as in 7A. Lower panel: Input samples are shown. Representative gel with control samples (C1, C2) and FAD samples (FAD1, FAD2) expressing mutants A260V and P264L respectively is shown. β-actin: loading control. Right: Graph shows relative levels of VEGFR2 co-precipitated with endoglin. (C), Brain sections from control and PS1 FAD patients were prepared as in Methods and stained for Col IV as in 1A. Upper: Representative images show brain vessels in either PS1 FAD or control (CT) brain sections. Scale bar: 80μm. Lower: Graph shows total vessel length density in PS1 FAD and CT brains quantified with Imaris software as in 1A. A-C , bars represent Mean ± S.E. For statistical analysis, unpaired t-test was performed. *p < 0.05, **p<0.01 and ***p<0.001.

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A), Brain tissue extracts were prepared as in Methods from twelve PS1 FAD patients each carrying a different PS1 mutation, and twelve non-demented controls described in Methods. Left: VEGFR2 dimers and monomers were detected in brain extracts on WBs using anti-VEGFR2 antibody D5B1. Representative gels with control (C1-4) or FAD samples (FAD1-4) expressing mutants P264L, A260V, N135S and P242H respectively are shown. Vinculin: loading control. Right: Graph shows the fold change in VEGFR2 dimer to monomer ratio of FAD and control samples. (B), Brain tissue extract from control and PS1 FAD patient brains described in 7A were prepared and IPed with anti-endoglin antibody (ab252345) or IgG as in Methods. Upper panel: VEGFR2 co-IPed with endoglin was detected on WBs using an anti-VEGFR2 antibody as in 7A. Lower panel: Input samples are shown. Representative gel with control samples (C1, C2) and FAD samples (FAD1, FAD2) expressing mutants A260V and P264L respectively is shown. β-actin: loading control. Right: Graph shows relative levels of VEGFR2 co-precipitated with endoglin. (C), Brain sections from control and PS1 FAD patients were prepared as in Methods and stained for Col IV as in 1A. Upper: Representative images show brain vessels in either PS1 FAD or control (CT) brain sections. Scale bar: 80μm. Lower: Graph shows total vessel length density in PS1 FAD and CT brains quantified with Imaris software as in 1A. A-C , bars represent Mean ± S.E. For statistical analysis, unpaired t-test was performed. *p < 0.05, **p<0.01 and ***p<0.001.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Mutagenesis, Control, Expressing, Staining, Software

Re suppresses ox-LDL-induced HUVEC proliferation and migration. (A, B) Results of the CCK-8 assay, n = 6. (C) Representative images of HUVECs at 0 h and 12 h after ox-LDL induction in wound healing experiments, bar = 50 μm. (D) Quantification of EC migration in the wound healing assay, n = 3. (E) Western blot assay and quantitative data of VE-cadherin and VEGFR2 in HUVECs, n = 3. ### p < 0.001, ## p < 0.01, # p < 0.05, vs. control group, ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05, vs. ox-LDL group.

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Re regulates PFKFB3-mediated glycolysis to inhibit endothelial cell migration to ameliorate atherosclerosis

doi: 10.1016/j.jgr.2025.11.012

Figure Lengend Snippet: Re suppresses ox-LDL-induced HUVEC proliferation and migration. (A, B) Results of the CCK-8 assay, n = 6. (C) Representative images of HUVECs at 0 h and 12 h after ox-LDL induction in wound healing experiments, bar = 50 μm. (D) Quantification of EC migration in the wound healing assay, n = 3. (E) Western blot assay and quantitative data of VE-cadherin and VEGFR2 in HUVECs, n = 3. ### p < 0.001, ## p < 0.01, # p < 0.05, vs. control group, ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05, vs. ox-LDL group.

Article Snippet: Ginsenoside Re, (Cat. B21055 , Shanghai Yuanye Biotechnology Co., Ltd, purity≥ 98 %); Simvastatin tablets (Cat. 20210925, Shandong Xinqi Pharmaceutical Co., Ltd); PFKFB3 antibody (Cat. D7H4Q, Cell Signaling Technology, Inc.); HIF-1α Rabbit pAb (Cat. A11945, ABclonal Biotechnology Co., Ltd); HK2 polyclonal antibody (Cat. 22029-1-AP, Wuhan Sanying Biotechnology Co., Ltd); vascular endothelial cadherin (VE-cadherin) antibody (Cat. 2158S, Cell Signaling Technology, Inc.); VEGFA Rabbit mAb (Cat. ab214424, abcam); VEGFR2 Rabbit mAb (Cat. 2479, Cell Signaling Technology, Inc.).

Techniques: Migration, CCK-8 Assay, Wound Healing Assay, Western Blot, Control

Re inhibits endothelial cell migration via the PFKFB3-HIF-1α/VEGFA-VEGFR2 signaling pathways. (A) Western blot assay and quantitative analysis of PFKFB3 in HUVECs, n = 3. (B) Cell viability of PFKFB3 overexpressing ECs measured by CCK-8 assay, n = 6. (C) Western blot assay and quantitative analysis of PFKFB3 in HUVECs, n = 3. (D–E) Representative images of HUVEC induced by ox-LDL for 0 h and 12 h in wound healing experiments (bar = 50 μm) and quantification of EC migration, n = 3. (F, G) Western blot assay and quantitative data of VE-cadherin, HIF-1α, VEGFA, and VEGFR2 in HUVECs, n = 3. ### p < 0.001, ## p < 0.01, # p < 0.05, vs. control group; ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05, vs. ox-LDL group.

Journal: Journal of Ginseng Research

Article Title: Ginsenoside Re regulates PFKFB3-mediated glycolysis to inhibit endothelial cell migration to ameliorate atherosclerosis

doi: 10.1016/j.jgr.2025.11.012

Figure Lengend Snippet: Re inhibits endothelial cell migration via the PFKFB3-HIF-1α/VEGFA-VEGFR2 signaling pathways. (A) Western blot assay and quantitative analysis of PFKFB3 in HUVECs, n = 3. (B) Cell viability of PFKFB3 overexpressing ECs measured by CCK-8 assay, n = 6. (C) Western blot assay and quantitative analysis of PFKFB3 in HUVECs, n = 3. (D–E) Representative images of HUVEC induced by ox-LDL for 0 h and 12 h in wound healing experiments (bar = 50 μm) and quantification of EC migration, n = 3. (F, G) Western blot assay and quantitative data of VE-cadherin, HIF-1α, VEGFA, and VEGFR2 in HUVECs, n = 3. ### p < 0.001, ## p < 0.01, # p < 0.05, vs. control group; ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05, vs. ox-LDL group.

Article Snippet: Ginsenoside Re, (Cat. B21055 , Shanghai Yuanye Biotechnology Co., Ltd, purity≥ 98 %); Simvastatin tablets (Cat. 20210925, Shandong Xinqi Pharmaceutical Co., Ltd); PFKFB3 antibody (Cat. D7H4Q, Cell Signaling Technology, Inc.); HIF-1α Rabbit pAb (Cat. A11945, ABclonal Biotechnology Co., Ltd); HK2 polyclonal antibody (Cat. 22029-1-AP, Wuhan Sanying Biotechnology Co., Ltd); vascular endothelial cadherin (VE-cadherin) antibody (Cat. 2158S, Cell Signaling Technology, Inc.); VEGFA Rabbit mAb (Cat. ab214424, abcam); VEGFR2 Rabbit mAb (Cat. 2479, Cell Signaling Technology, Inc.).

Techniques: Migration, Protein-Protein interactions, Western Blot, CCK-8 Assay, Control

(A) HUVECs were infected with Ad-Moesin WT or Ad-Moesin T558D for 48 hrs, and total RNA was harvested for RNA-seq. The differentially expressed genes were analyzed with Gene Ontology Enrichment Analysis. The top nine gene sets are shown. Data were analyzed from GSE302986. (B) HUVECs transfected with siCtrl or si KINDLIN-2 were serum-starved (2% FBS) for 8 hrs, then stimulated with 2 μg/mL Alexa 594-labeled VEGF (red) for 30 mins. VEGF accumulation was visualized by confocal microscopy. Actin, phalloidin (green); nuclei, DAPI (blue). (C) Quantification of Alexa 594-labeled VEGF spots normalized to cellular area (n= 4 independent experiments). (D) HUVECs infected with Ad-Moesin WT or Ad-Moesin T558D were starved and stimulated with Alexa 594-labeled VEGF as in (B). VEGF accumulation was imaged by confocal microscopy. (E) Quantification of Alexa 594-labeled VEGF spots normalized to cellular area (n=3 independent experiments). (F) HUVECs transfected with siCtrl or si KINDLIN-2 were serum-starved for 8 hrs, stimulated with 50 ng/ml VEGF for 30 mins, fixed, and stained for VEGFR2 (green). Representative images show intracellular VEGFR2 vesicles. (G) Quantification of VEGFR2 vesicle number per cell from (F) (n=3 independent experiments). (H) HUVECs infected with Ad-Moesin WT or Ad-Moesin T558D were treated as in (F), and stained for VEGFR2. (I) Quantification of VEGFR2 vesicle number per cell from (H) (n= 3 independent experiments). (J) Schematic of the cell surface biotinylation assay. HUVECs were starved for 8 hrs, labeled with EZ-Link Sulfo-NHS-SS-Biotin (0.25 mg/mL) at 4 °C for 1 hr, then stimulated with VEGF (50 ng/mL) for 30 mins. After surface biotin was stripped with GSH elution buffer, total proteins were extracted. Biotinylated internalized VEGFR2 was pulled down using streptavidin magnetic beads and analyzed by western blotting. (K) Cell surface biotinylation assay for VEGFR2 internalization in siCtrl- and si KINDLIN-2 –transfected HUVECs. Input lysates show VEGFR2, Kindlin-2, and GAPDH. “Surf” represents surface VEGFR2 prior to VEGF stimulation and biotin stripping. (L) Quantification of internalized VEGFR2 normalized to surface VEGFR2 levels (n=4 independent experiments). (M) Schematic of in vivo Alexa 594-labeled VEGF uptake assay in Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. Tamoxifen was administered from P1 to P3; Alexa 594-labeled VEGF was injected intravitreally at P6 and analyzed after 30 mins. (N) Representative confocal images of the retinal angiogenic front showing uptake of Alexa 594-labeled VEGF by ECs from Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. (O) Quantification of internalized Alexa 594-VEGF at the angiogenic front (n = 4/4 pups). (P) Western blot of HUVECs transfected with siCtrl or si KINDLIN-2 , starved for 8 hrs, then stimulated with 50 ng/mL VEGF. (Q) Quantification of p-ERK levels at 5 mins after VEGF stimulation from (P) (n = 4 independent experiments). (R) Western blot of HUVECs infected with Ad-Vector, Ad-Moesin WT , or Ad-Moesin T558D , treated as in (P). (S) Quantification of p-ERK levels at 5 mins after VEGF stimulation from (R) (n = 3 independent experiments). Data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 by two-tailed Student’s t-test or one-way ANOVA followed by Tukey’s multiple comparisons test. Scale bars: 5 μm in (B), (D), (F), (H), and (N).

Journal: bioRxiv

Article Title: Kindlin-2-Moesin interaction orchestrates sprouting angiogenesis via modulating endothelial membrane mechanics and VEGF signaling

doi: 10.64898/2026.02.24.707842

Figure Lengend Snippet: (A) HUVECs were infected with Ad-Moesin WT or Ad-Moesin T558D for 48 hrs, and total RNA was harvested for RNA-seq. The differentially expressed genes were analyzed with Gene Ontology Enrichment Analysis. The top nine gene sets are shown. Data were analyzed from GSE302986. (B) HUVECs transfected with siCtrl or si KINDLIN-2 were serum-starved (2% FBS) for 8 hrs, then stimulated with 2 μg/mL Alexa 594-labeled VEGF (red) for 30 mins. VEGF accumulation was visualized by confocal microscopy. Actin, phalloidin (green); nuclei, DAPI (blue). (C) Quantification of Alexa 594-labeled VEGF spots normalized to cellular area (n= 4 independent experiments). (D) HUVECs infected with Ad-Moesin WT or Ad-Moesin T558D were starved and stimulated with Alexa 594-labeled VEGF as in (B). VEGF accumulation was imaged by confocal microscopy. (E) Quantification of Alexa 594-labeled VEGF spots normalized to cellular area (n=3 independent experiments). (F) HUVECs transfected with siCtrl or si KINDLIN-2 were serum-starved for 8 hrs, stimulated with 50 ng/ml VEGF for 30 mins, fixed, and stained for VEGFR2 (green). Representative images show intracellular VEGFR2 vesicles. (G) Quantification of VEGFR2 vesicle number per cell from (F) (n=3 independent experiments). (H) HUVECs infected with Ad-Moesin WT or Ad-Moesin T558D were treated as in (F), and stained for VEGFR2. (I) Quantification of VEGFR2 vesicle number per cell from (H) (n= 3 independent experiments). (J) Schematic of the cell surface biotinylation assay. HUVECs were starved for 8 hrs, labeled with EZ-Link Sulfo-NHS-SS-Biotin (0.25 mg/mL) at 4 °C for 1 hr, then stimulated with VEGF (50 ng/mL) for 30 mins. After surface biotin was stripped with GSH elution buffer, total proteins were extracted. Biotinylated internalized VEGFR2 was pulled down using streptavidin magnetic beads and analyzed by western blotting. (K) Cell surface biotinylation assay for VEGFR2 internalization in siCtrl- and si KINDLIN-2 –transfected HUVECs. Input lysates show VEGFR2, Kindlin-2, and GAPDH. “Surf” represents surface VEGFR2 prior to VEGF stimulation and biotin stripping. (L) Quantification of internalized VEGFR2 normalized to surface VEGFR2 levels (n=4 independent experiments). (M) Schematic of in vivo Alexa 594-labeled VEGF uptake assay in Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. Tamoxifen was administered from P1 to P3; Alexa 594-labeled VEGF was injected intravitreally at P6 and analyzed after 30 mins. (N) Representative confocal images of the retinal angiogenic front showing uptake of Alexa 594-labeled VEGF by ECs from Kindlin-2 WT and Kindlin-2 iΔEC(Pdgfb) pups. (O) Quantification of internalized Alexa 594-VEGF at the angiogenic front (n = 4/4 pups). (P) Western blot of HUVECs transfected with siCtrl or si KINDLIN-2 , starved for 8 hrs, then stimulated with 50 ng/mL VEGF. (Q) Quantification of p-ERK levels at 5 mins after VEGF stimulation from (P) (n = 4 independent experiments). (R) Western blot of HUVECs infected with Ad-Vector, Ad-Moesin WT , or Ad-Moesin T558D , treated as in (P). (S) Quantification of p-ERK levels at 5 mins after VEGF stimulation from (R) (n = 3 independent experiments). Data are presented as mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 by two-tailed Student’s t-test or one-way ANOVA followed by Tukey’s multiple comparisons test. Scale bars: 5 μm in (B), (D), (F), (H), and (N).

Article Snippet: Samples were then blocked in PBS containing 2% BSA and 0.1% Triton X-100 for 1 hr at room temperature, followed by staining with primary antibody VEGFR2 (Cell Signaling Technology, #2479S, 1:200) and the appropriate secondary antibody.

Techniques: Infection, RNA Sequencing, Transfection, Labeling, Confocal Microscopy, Staining, Cell Surface Biotinylation Assay, Magnetic Beads, Western Blot, Stripping Membranes, In Vivo, Injection, Plasmid Preparation, Two Tailed Test

(A) Volcano plot of differentially secreted proteins from primary LSECs treated with 40 ng/mL LIF versus control (n = 4). (B&C) qPCR of lifr (B) and hgf (C) levels in LSECs isolated from lifr f/f mice with indicated treatments. (D) HGF concentrations in conditioned medium from LSECs treated with hLIF at the indicated doses. (E) HGF concentrations in conditioned medium from LSECs treated with 40 ng/mL hLIF in the presence of the STAT3 inhibitor C188-9 at the indicated doses. (F) HGF concentrations in conditioned medium from LSECs isolated from lifr f/f or lifrΔEC mice treated with hLIF, hCT-1 or hOSM. (G) Western blot analysis of hepatocytes co-cultured with LSECs with or without 40 ng/mL LIF. Quantification is shown in Figure S4C. (H) Model depicting LIF action on LSECs to stimulate angiogenesis and HGF release, thereby supporting hepatocyte proliferation via paracrine regulation.

Journal: bioRxiv

Article Title: The LIF-LIFR Axis Promotes Liver Regeneration via Modulation of Angiogenesis and HGF Release from LSECs

doi: 10.64898/2026.02.24.707802

Figure Lengend Snippet: (A) Volcano plot of differentially secreted proteins from primary LSECs treated with 40 ng/mL LIF versus control (n = 4). (B&C) qPCR of lifr (B) and hgf (C) levels in LSECs isolated from lifr f/f mice with indicated treatments. (D) HGF concentrations in conditioned medium from LSECs treated with hLIF at the indicated doses. (E) HGF concentrations in conditioned medium from LSECs treated with 40 ng/mL hLIF in the presence of the STAT3 inhibitor C188-9 at the indicated doses. (F) HGF concentrations in conditioned medium from LSECs isolated from lifr f/f or lifrΔEC mice treated with hLIF, hCT-1 or hOSM. (G) Western blot analysis of hepatocytes co-cultured with LSECs with or without 40 ng/mL LIF. Quantification is shown in Figure S4C. (H) Model depicting LIF action on LSECs to stimulate angiogenesis and HGF release, thereby supporting hepatocyte proliferation via paracrine regulation.

Article Snippet: The following primary antibodies were used in this study : LIFR (Proteintech, 22779-1-AP, 1:1,000 dilution), Tubulin (Proteintech, 66031-1-Ig, 1:2,000 dilution), Actin (Proteintech, 66009-1-Ig, 1:10,000 dilution), PCNA (Abcam, AB220208, 1:2,000 dilution), p-Y1175-VEGFR2 (Cell Signaling Technology, 3770S, 1:2000 dilution), Total-VEGFR2 (Cell Signaling Technology, 2479S, 1:2000 dilution), p-T705-STAT3 (Cell Signaling Technology, 9145S, 1:2000 dilution), Total-STAT3 (Cell Signaling Technology, 30835S, 1:2,000 dilution), p-T202/Y204-ERK (Cell Signaling Technology, 4370, 1:2,000 dilution) and Total-ERK (Cell Signaling Technology, 4695, 1:2,000 dilution), p-S473-AKT (Cell Signaling Technology, 4060, 1:2,000 dilution), Total-AKT (Cell Signaling Technology, 2920, 1:2000 dilution)

Techniques: Control, Isolation, Western Blot, Cell Culture