placental pericytes Search Results


core spheroids  (Angio-Proteomie)
94
Angio-Proteomie core spheroids
Core Spheroids, supplied by Angio-Proteomie, 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/placental+pericytes/Human+Placental+Microvascular+Pericytes/10__21203_slash_rs__3__rs___1144566_slash_v1-397-7-17
Average 94 stars, based on 1 article reviews
core spheroids - by Bioz Stars, 2026-10
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red fluorescent protein rfp labeled huvecs  (Angio-Proteomie)
90
Angio-Proteomie red fluorescent protein rfp labeled huvecs
(a) Side and top views of the culture dish before setup. The dish was a normal 12-phi glass-bottom dish. A glass separator was set at the center of the dish with a bioinert adhesive. (b) First, we placed 150 μl fibrin gel mixed with <t>HUVECs</t> in the center of the glass plate, so that two sides of the dish were separated by the glass separator and fibrin gel. We also added LF-containing fibrin gel at the edge of the dish. (c) We incubated the dish for 30 min to solidify the fibrin gel. (d) We added 1 ml culture medium to both wells and incubated the dish for 1 week. (e) After 1 week of culture, a vascular network with a perfusable lumen was formed in the glass-bottom region. Then, we cut both edges of the regions to make openings. (f) After the cuts, we increased the amount of culture medium on one side of the dish. This caused a static pressure difference between both openings of the self-organized capillary network, resulting in steady flow inside the apparatus. (g) Low magnification view of the self-organized vascular network. RFP-HUVECs were cultivated in the fibrin gel, and we observed the vascular network formation. (h) High magnification view of (g). Vascular network with a lumen was generated in the fibrin gel region. (i) Low magnification view of the self-organized vascular network after long-term culture with flow. (j) Visualization of the perfusable area by FITC-dextran. Perfusable regions existed near the inlet and outlet, near the glass separator and edge of the well. (k) Flow inside the lumen was visualized using <t>fluorescent</t> beads. (l) Snapshot of the culture system when using whole blood as a tracer. Red blood cells were flowing inside the self-organized vasculature. (m) Projection of multiple frames of (l). Movements of red blood cells were visualized as a stream. Scale bars: 3 mm (g, i, j); 100 μm (h); 500 μm (k); 50 μm (l, m).
Red Fluorescent Protein Rfp Labeled Huvecs, supplied by Angio-Proteomie, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/placental+pericytes/GFP+Expressing+Human+Placental+Microvascular+Pericytes/pmc07592787-39-5-18
Average 90 stars, based on 1 article reviews
red fluorescent protein rfp labeled huvecs - by Bioz Stars, 2026-10
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placental-derived primary human pericytes #per-f  (ZenBio)
90
ZenBio placental-derived primary human pericytes #per-f
PMA actively downregulates TF in primary human <t>pericytes.</t> Pericyte cultures were treated with PMA for the indicated times. The mean +/− standard deviation (SD) of 3 independent experiments is shown. *p<0.05, **p<0.01, ***p<0.001. A. Expression of TF protein was assessed by western blot. B. For each time point the relative amount of TF protein expressed by PMA-treated cells is shown as a percentage of TF expressed by vehicle-treated cells. C. Activity of total TF as measured in lysed pericytes was determined by measuring cleavage of a FXa-specific chromogenic substrate as described in “Methods”. TF activity is presented as the rate of FXa generation. D. Expression of TF mRNA was quantitated by qRT-PCR. For each time point the amount of TF mRNA expressed by PMA-treated cells is presented as the fold change relative to TF mRNA expressed by vehicle-treated control cells.
Placental Derived Primary Human Pericytes #Per F, supplied by ZenBio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/placental+pericytes/placental+derived+primary+human+pericytes++per+f/pmc06813842-129-2-13
Average 90 stars, based on 1 article reviews
placental-derived primary human pericytes #per-f - by Bioz Stars, 2026-10
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gfp-human placental microvascular pericytes  (AcceGen Biotechnology)
N/A
GFP-HPlMVPCs are Puromycin resistant HPlMVPCs isolated from normal human placental tissue, after infected GFP expressing lentiviral particles. The cells are shipped in frozen vials (the cells are provided @ passage 3). Pericyte Growth Medium is
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Image Search Results


(a) Side and top views of the culture dish before setup. The dish was a normal 12-phi glass-bottom dish. A glass separator was set at the center of the dish with a bioinert adhesive. (b) First, we placed 150 μl fibrin gel mixed with HUVECs in the center of the glass plate, so that two sides of the dish were separated by the glass separator and fibrin gel. We also added LF-containing fibrin gel at the edge of the dish. (c) We incubated the dish for 30 min to solidify the fibrin gel. (d) We added 1 ml culture medium to both wells and incubated the dish for 1 week. (e) After 1 week of culture, a vascular network with a perfusable lumen was formed in the glass-bottom region. Then, we cut both edges of the regions to make openings. (f) After the cuts, we increased the amount of culture medium on one side of the dish. This caused a static pressure difference between both openings of the self-organized capillary network, resulting in steady flow inside the apparatus. (g) Low magnification view of the self-organized vascular network. RFP-HUVECs were cultivated in the fibrin gel, and we observed the vascular network formation. (h) High magnification view of (g). Vascular network with a lumen was generated in the fibrin gel region. (i) Low magnification view of the self-organized vascular network after long-term culture with flow. (j) Visualization of the perfusable area by FITC-dextran. Perfusable regions existed near the inlet and outlet, near the glass separator and edge of the well. (k) Flow inside the lumen was visualized using fluorescent beads. (l) Snapshot of the culture system when using whole blood as a tracer. Red blood cells were flowing inside the self-organized vasculature. (m) Projection of multiple frames of (l). Movements of red blood cells were visualized as a stream. Scale bars: 3 mm (g, i, j); 100 μm (h); 500 μm (k); 50 μm (l, m).

Journal: PLoS ONE

Article Title: A new perfusion culture method with a self-organized capillary network

doi: 10.1371/journal.pone.0240552

Figure Lengend Snippet: (a) Side and top views of the culture dish before setup. The dish was a normal 12-phi glass-bottom dish. A glass separator was set at the center of the dish with a bioinert adhesive. (b) First, we placed 150 μl fibrin gel mixed with HUVECs in the center of the glass plate, so that two sides of the dish were separated by the glass separator and fibrin gel. We also added LF-containing fibrin gel at the edge of the dish. (c) We incubated the dish for 30 min to solidify the fibrin gel. (d) We added 1 ml culture medium to both wells and incubated the dish for 1 week. (e) After 1 week of culture, a vascular network with a perfusable lumen was formed in the glass-bottom region. Then, we cut both edges of the regions to make openings. (f) After the cuts, we increased the amount of culture medium on one side of the dish. This caused a static pressure difference between both openings of the self-organized capillary network, resulting in steady flow inside the apparatus. (g) Low magnification view of the self-organized vascular network. RFP-HUVECs were cultivated in the fibrin gel, and we observed the vascular network formation. (h) High magnification view of (g). Vascular network with a lumen was generated in the fibrin gel region. (i) Low magnification view of the self-organized vascular network after long-term culture with flow. (j) Visualization of the perfusable area by FITC-dextran. Perfusable regions existed near the inlet and outlet, near the glass separator and edge of the well. (k) Flow inside the lumen was visualized using fluorescent beads. (l) Snapshot of the culture system when using whole blood as a tracer. Red blood cells were flowing inside the self-organized vasculature. (m) Projection of multiple frames of (l). Movements of red blood cells were visualized as a stream. Scale bars: 3 mm (g, i, j); 100 μm (h); 500 μm (k); 50 μm (l, m).

Article Snippet: For visualization purposes, we used red fluorescent protein (RFP)-labeled HUVECs and GFP-labeled human placental microvascular pericytes (cAP-0029GFP) from Angio-proteomie Inc. HL60 and NMuMG-Fucci cells were provided by the Riken Bioresource Research Center (RCB2813 and RCB0041, respectively).

Techniques: Adhesive, Incubation, Generated

(a) Experimental procedure. Spheroids containing RFP-HUVECs and lung fibroblast were generated and embedded in fibrin gel. After 1 week, sprouts from the spheroids became sufficiently long. Then, we cut the tip of the sprouts from both sides of the well and exerted static pressure to one side of the well. (b) Visualization of the perfusion inside the spheroid using FITC-dextran. (c) High-magnification view of the sprouts connecting two spheroids. FITC-dextran is running through the sprout structure. Scale bar: 1 mm (b); 100 μm (c).

Journal: PLoS ONE

Article Title: A new perfusion culture method with a self-organized capillary network

doi: 10.1371/journal.pone.0240552

Figure Lengend Snippet: (a) Experimental procedure. Spheroids containing RFP-HUVECs and lung fibroblast were generated and embedded in fibrin gel. After 1 week, sprouts from the spheroids became sufficiently long. Then, we cut the tip of the sprouts from both sides of the well and exerted static pressure to one side of the well. (b) Visualization of the perfusion inside the spheroid using FITC-dextran. (c) High-magnification view of the sprouts connecting two spheroids. FITC-dextran is running through the sprout structure. Scale bar: 1 mm (b); 100 μm (c).

Article Snippet: For visualization purposes, we used red fluorescent protein (RFP)-labeled HUVECs and GFP-labeled human placental microvascular pericytes (cAP-0029GFP) from Angio-proteomie Inc. HL60 and NMuMG-Fucci cells were provided by the Riken Bioresource Research Center (RCB2813 and RCB0041, respectively).

Techniques: Generated

(a) Initial shape of the vascular network. HUVECs were stained with UEA1 and nuclei were stained with Hoechst 33342. There was a flow-positive region (yellow-dashed line) and non-flow region (green-dashed line). Direction of flow is indicated by a white allow. (b) Kymograph of the yellow-dashed line region. Horizontal direction represents space, and vertical direction represents time. Collective movement toward upstream of the flow was observed. White arrow indicates the flow of cell debris. (c) Kymograph of the green-dashed line region. Cell movement was random. (d–n) Cell shape changes induced by flow: (d, e) Fast flow. When FITC dextran was perfused, the vessel regions near the inlet or outlet showed fast flow. (f) Brightfield view of the fast flow region. Endothelial cells became shaped as spindles aligned parallel to the flow direction. (g) Fluorescence view of the fast flow region. At the floor of the lumen, we observed spindle-shaped cells parallel to the flow direction. (h, i) Slow flow. When FITC dextran was perfused, the vessel regions far from the inlet or outlet showed slow flow. (j) Brightfield view and (k) confocal view of the slow flow region. Endothelial cells did not show any polarity. (l) Low magnification view of the non-flow region. (m) Brightfield view and (n) confocal view of the non-flow region. Vasculatures were disconnected and became thin endothelial cysts with cell debris inside. Scale bars: 1 mm (d, e, h, i, l); 200 μm (f, g, j, k, m, n).

Journal: PLoS ONE

Article Title: A new perfusion culture method with a self-organized capillary network

doi: 10.1371/journal.pone.0240552

Figure Lengend Snippet: (a) Initial shape of the vascular network. HUVECs were stained with UEA1 and nuclei were stained with Hoechst 33342. There was a flow-positive region (yellow-dashed line) and non-flow region (green-dashed line). Direction of flow is indicated by a white allow. (b) Kymograph of the yellow-dashed line region. Horizontal direction represents space, and vertical direction represents time. Collective movement toward upstream of the flow was observed. White arrow indicates the flow of cell debris. (c) Kymograph of the green-dashed line region. Cell movement was random. (d–n) Cell shape changes induced by flow: (d, e) Fast flow. When FITC dextran was perfused, the vessel regions near the inlet or outlet showed fast flow. (f) Brightfield view of the fast flow region. Endothelial cells became shaped as spindles aligned parallel to the flow direction. (g) Fluorescence view of the fast flow region. At the floor of the lumen, we observed spindle-shaped cells parallel to the flow direction. (h, i) Slow flow. When FITC dextran was perfused, the vessel regions far from the inlet or outlet showed slow flow. (j) Brightfield view and (k) confocal view of the slow flow region. Endothelial cells did not show any polarity. (l) Low magnification view of the non-flow region. (m) Brightfield view and (n) confocal view of the non-flow region. Vasculatures were disconnected and became thin endothelial cysts with cell debris inside. Scale bars: 1 mm (d, e, h, i, l); 200 μm (f, g, j, k, m, n).

Article Snippet: For visualization purposes, we used red fluorescent protein (RFP)-labeled HUVECs and GFP-labeled human placental microvascular pericytes (cAP-0029GFP) from Angio-proteomie Inc. HL60 and NMuMG-Fucci cells were provided by the Riken Bioresource Research Center (RCB2813 and RCB0041, respectively).

Techniques: Staining, Fluorescence

(a) Experiment setup. We prepared two groups of dishes in which a mixture of RFP-HUVECs and NMuMG-Fucci cells were seeded in the fibrin gel. After the perfusable network was formed, we made openings to both groups but transferred medium in only one group of the dishes. (b) Typical appearance of the NMuMG cell colony and its schematic representation. NMuMG-Fucci cells (red and green nuclei) forms colonies outside the vascular lumen. (c) Time course of the cell division monitored by Fucci reporter. (d) Time course of GFP(+) area ratio. 20 NMuMG-Fucci colonies were observed, and the area of GFP(+) areas was obtained using Fiji. (e) GFP(+) area ratio between day 1 and day 0. Statistically significant difference was detected (Student t-test). Scale bars: 50 μm.

Journal: PLoS ONE

Article Title: A new perfusion culture method with a self-organized capillary network

doi: 10.1371/journal.pone.0240552

Figure Lengend Snippet: (a) Experiment setup. We prepared two groups of dishes in which a mixture of RFP-HUVECs and NMuMG-Fucci cells were seeded in the fibrin gel. After the perfusable network was formed, we made openings to both groups but transferred medium in only one group of the dishes. (b) Typical appearance of the NMuMG cell colony and its schematic representation. NMuMG-Fucci cells (red and green nuclei) forms colonies outside the vascular lumen. (c) Time course of the cell division monitored by Fucci reporter. (d) Time course of GFP(+) area ratio. 20 NMuMG-Fucci colonies were observed, and the area of GFP(+) areas was obtained using Fiji. (e) GFP(+) area ratio between day 1 and day 0. Statistically significant difference was detected (Student t-test). Scale bars: 50 μm.

Article Snippet: For visualization purposes, we used red fluorescent protein (RFP)-labeled HUVECs and GFP-labeled human placental microvascular pericytes (cAP-0029GFP) from Angio-proteomie Inc. HL60 and NMuMG-Fucci cells were provided by the Riken Bioresource Research Center (RCB2813 and RCB0041, respectively).

Techniques:

(a) LM4-GFP cells were introduced into the self-organized capillary network consisting of HUVECs visualized by UEA-1 lectin. (b) High magnification time-lapse view of (a). We observed cancer cells emigrating out of the blood vessels. (c) Detailed morphology of cancer cells on the endothelial cells. Emigrated LM4 cells attached to the blood vessel with highly polarized morphology and multiple protrusions. (c) Max projection image, (c') orthogonal section, and (c'') 3D-reconstructed image. White arrows: cancer cell protrusions. Scale bars: 50 μm (a); 10 μm (b, c).

Journal: PLoS ONE

Article Title: A new perfusion culture method with a self-organized capillary network

doi: 10.1371/journal.pone.0240552

Figure Lengend Snippet: (a) LM4-GFP cells were introduced into the self-organized capillary network consisting of HUVECs visualized by UEA-1 lectin. (b) High magnification time-lapse view of (a). We observed cancer cells emigrating out of the blood vessels. (c) Detailed morphology of cancer cells on the endothelial cells. Emigrated LM4 cells attached to the blood vessel with highly polarized morphology and multiple protrusions. (c) Max projection image, (c') orthogonal section, and (c'') 3D-reconstructed image. White arrows: cancer cell protrusions. Scale bars: 50 μm (a); 10 μm (b, c).

Article Snippet: For visualization purposes, we used red fluorescent protein (RFP)-labeled HUVECs and GFP-labeled human placental microvascular pericytes (cAP-0029GFP) from Angio-proteomie Inc. HL60 and NMuMG-Fucci cells were provided by the Riken Bioresource Research Center (RCB2813 and RCB0041, respectively).

Techniques:

PMA actively downregulates TF in primary human pericytes. Pericyte cultures were treated with PMA for the indicated times. The mean +/− standard deviation (SD) of 3 independent experiments is shown. *p<0.05, **p<0.01, ***p<0.001. A. Expression of TF protein was assessed by western blot. B. For each time point the relative amount of TF protein expressed by PMA-treated cells is shown as a percentage of TF expressed by vehicle-treated cells. C. Activity of total TF as measured in lysed pericytes was determined by measuring cleavage of a FXa-specific chromogenic substrate as described in “Methods”. TF activity is presented as the rate of FXa generation. D. Expression of TF mRNA was quantitated by qRT-PCR. For each time point the amount of TF mRNA expressed by PMA-treated cells is presented as the fold change relative to TF mRNA expressed by vehicle-treated control cells.

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: PMA actively downregulates TF in primary human pericytes. Pericyte cultures were treated with PMA for the indicated times. The mean +/− standard deviation (SD) of 3 independent experiments is shown. *p<0.05, **p<0.01, ***p<0.001. A. Expression of TF protein was assessed by western blot. B. For each time point the relative amount of TF protein expressed by PMA-treated cells is shown as a percentage of TF expressed by vehicle-treated cells. C. Activity of total TF as measured in lysed pericytes was determined by measuring cleavage of a FXa-specific chromogenic substrate as described in “Methods”. TF activity is presented as the rate of FXa generation. D. Expression of TF mRNA was quantitated by qRT-PCR. For each time point the amount of TF mRNA expressed by PMA-treated cells is presented as the fold change relative to TF mRNA expressed by vehicle-treated control cells.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Standard Deviation, Expressing, Western Blot, Activity Assay, Quantitative RT-PCR, Control

Internalized TF is degraded primarily in lysosomes. Pericytes were treated with MG132 or chloroquine prior to PMA for the indicated times. The mean + SD of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Relative amounts of internalized TF and expression of total TF were assessed by biotin labeling of cell surface proteins and western blotting as in “Methods”. B. The amounts of internalized TF are expressed as percentages relative to the total amount of TF expressed on the cell surface. C. Relative amounts of total TF expressed by PMA-treated pericytes are shown as percentages of TF expressed by control cells. D. Pericytes were stained for TF (green), the lysosomal marker LAMP-1 (red), and nuclei (blue, DAPI). E. Co-localization of TF and LAMP-1 was determined by calculating Pearson’s Correlation Coefficient with Image J, Fiji Plugin.

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: Internalized TF is degraded primarily in lysosomes. Pericytes were treated with MG132 or chloroquine prior to PMA for the indicated times. The mean + SD of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Relative amounts of internalized TF and expression of total TF were assessed by biotin labeling of cell surface proteins and western blotting as in “Methods”. B. The amounts of internalized TF are expressed as percentages relative to the total amount of TF expressed on the cell surface. C. Relative amounts of total TF expressed by PMA-treated pericytes are shown as percentages of TF expressed by control cells. D. Pericytes were stained for TF (green), the lysosomal marker LAMP-1 (red), and nuclei (blue, DAPI). E. Co-localization of TF and LAMP-1 was determined by calculating Pearson’s Correlation Coefficient with Image J, Fiji Plugin.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Expressing, Labeling, Western Blot, Control, Staining, Marker

PMA shortens the half-life of TF protein while leaving degradation of TF mRNA unaffected. The mean +/− standard deviation (SD) of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Pericytes were pre-treated with Cyclohexamide (CHX) prior to addition of vehicle or PMA for the indicated times. Degradation of TF protein was assessed by western blot. B. Relative amounts of TF protein expressed by CHX-treated cells are given as a percentage of TF expressed by non-CHX-treated control cells. C. Pericytes were pre-treated with Actinomycin D (ActD) prior to receiving vehicle or PMA for the indicated times. Degradation of TF mRNA was analyzed by qRT-PCR. The amount of TF mRNA expressed by ActD-treated pericytes relative to non-ActD-treated control cells is shown.

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: PMA shortens the half-life of TF protein while leaving degradation of TF mRNA unaffected. The mean +/− standard deviation (SD) of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Pericytes were pre-treated with Cyclohexamide (CHX) prior to addition of vehicle or PMA for the indicated times. Degradation of TF protein was assessed by western blot. B. Relative amounts of TF protein expressed by CHX-treated cells are given as a percentage of TF expressed by non-CHX-treated control cells. C. Pericytes were pre-treated with Actinomycin D (ActD) prior to receiving vehicle or PMA for the indicated times. Degradation of TF mRNA was analyzed by qRT-PCR. The amount of TF mRNA expressed by ActD-treated pericytes relative to non-ActD-treated control cells is shown.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Standard Deviation, Western Blot, Control, Quantitative RT-PCR

Inhibition of Protein Kinase C attenuates PMA-mediated downregulation of TF. Pericytes were pre-treated with Go 6983 or GFX prior to PMA for the indicated times. The mean + SD of 3 independent experiments is shown. *p<0.5, **p<0.1, ***p<0.001. A. TF protein expression was analyzed by western blot. B. For each time point the amount of TF protein is shown as a percentage of the TF expressed by control cells. C. Expression of TF mRNA was quantified by qRT-PCR. The amount of expressed TF mRNA is presented as the fold change relative to TF mRNA expressed by control cells. D. Activity of total TF in lysed pericytes is shown as the rate of FXa generation.

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: Inhibition of Protein Kinase C attenuates PMA-mediated downregulation of TF. Pericytes were pre-treated with Go 6983 or GFX prior to PMA for the indicated times. The mean + SD of 3 independent experiments is shown. *p<0.5, **p<0.1, ***p<0.001. A. TF protein expression was analyzed by western blot. B. For each time point the amount of TF protein is shown as a percentage of the TF expressed by control cells. C. Expression of TF mRNA was quantified by qRT-PCR. The amount of expressed TF mRNA is presented as the fold change relative to TF mRNA expressed by control cells. D. Activity of total TF in lysed pericytes is shown as the rate of FXa generation.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Inhibition, Expressing, Western Blot, Control, Quantitative RT-PCR, Activity Assay

PMA increases internalization of TF from the pericyte surface. Where specified, pericytes were pre-treated with either Go 6983 or GFX prior to receiving PMA for the times indicated. The mean +SD of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Pericyte cultures were fixed and stained for TF (green). Prominent punctate staining for TF was seen 4 hours after PMA while TF was virtually absent 8 hours after PMA. B. The total TF content of pericytes was determined by western blotting of whole cell lysates. Internalized surface TF was determined by labeling cell surface proteins with biotin, isolating biotin-labeled proteins from cell lysates, and assessing their TF content by western blotting. C. Relative amounts of internalized TF are shown as a percentage of the total surface TF. D. Relative amounts of total TF are presented as a percentage of the total surface TF expressed by vehicle-treated control cells. E. Activity of surface TF was determined on whole cells by measuring the rate of FXa generation.

Journal: Journal of thrombosis and haemostasis : JTH

Article Title: A Unique Protein Kinase C-Dependent Pathway for Tissue Factor Downregulation in Pericytes

doi: 10.1111/jth.14399

Figure Lengend Snippet: PMA increases internalization of TF from the pericyte surface. Where specified, pericytes were pre-treated with either Go 6983 or GFX prior to receiving PMA for the times indicated. The mean +SD of 3 independent experiments is shown. **p<0.01, ***p<0.001. A. Pericyte cultures were fixed and stained for TF (green). Prominent punctate staining for TF was seen 4 hours after PMA while TF was virtually absent 8 hours after PMA. B. The total TF content of pericytes was determined by western blotting of whole cell lysates. Internalized surface TF was determined by labeling cell surface proteins with biotin, isolating biotin-labeled proteins from cell lysates, and assessing their TF content by western blotting. C. Relative amounts of internalized TF are shown as a percentage of the total surface TF. D. Relative amounts of total TF are presented as a percentage of the total surface TF expressed by vehicle-treated control cells. E. Activity of surface TF was determined on whole cells by measuring the rate of FXa generation.

Article Snippet: Placental-derived primary human pericytes (#PER-F) and pericyte growth media (#PER-1) were purchased from Zen-bio, Inc. Pericytes were cultured according to manufacturer’s protocol.

Techniques: Staining, Western Blot, Labeling, Control, Activity Assay