Structured Review

Cusabio vegf a
Additional <t>VEGF-A</t> is required to recapitulate the phenotypes of DR. ( a ) Diagram depicting the experiment schedule for selective loss of pericytes in retinal vessels using adult DTA iΔPC mice, intra-vitreal administration of VEGF-A (1 μg) into one eye and BSA (1 μg) into the contralateral eye, and analyses at 4 days after the administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, PDGFRβ + pericyte coverage, expressions of Ang2 and VEGFR2 in CD31 + vessels, and dextran (70 kDa) leakage in retinas of WT and DTA iΔPC mice that were intra-vitreally treated with BSA (B) or VEGF-A (VA). Each group, n =6. Error bars represent mean±s.d. * P
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Images

1) Product Images from "Plastic roles of pericytes in the blood–retinal barrier"

Article Title: Plastic roles of pericytes in the blood–retinal barrier

Journal: Nature Communications

doi: 10.1038/ncomms15296

Additional VEGF-A is required to recapitulate the phenotypes of DR. ( a ) Diagram depicting the experiment schedule for selective loss of pericytes in retinal vessels using adult DTA iΔPC mice, intra-vitreal administration of VEGF-A (1 μg) into one eye and BSA (1 μg) into the contralateral eye, and analyses at 4 days after the administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, PDGFRβ + pericyte coverage, expressions of Ang2 and VEGFR2 in CD31 + vessels, and dextran (70 kDa) leakage in retinas of WT and DTA iΔPC mice that were intra-vitreally treated with BSA (B) or VEGF-A (VA). Each group, n =6. Error bars represent mean±s.d. * P
Figure Legend Snippet: Additional VEGF-A is required to recapitulate the phenotypes of DR. ( a ) Diagram depicting the experiment schedule for selective loss of pericytes in retinal vessels using adult DTA iΔPC mice, intra-vitreal administration of VEGF-A (1 μg) into one eye and BSA (1 μg) into the contralateral eye, and analyses at 4 days after the administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, PDGFRβ + pericyte coverage, expressions of Ang2 and VEGFR2 in CD31 + vessels, and dextran (70 kDa) leakage in retinas of WT and DTA iΔPC mice that were intra-vitreally treated with BSA (B) or VEGF-A (VA). Each group, n =6. Error bars represent mean±s.d. * P

Techniques Used: Mouse Assay

Tie2 activation lessens BRB disintegration in pericyte-free adult retina. ( a ) Diagram depicting the experiment schedule for selective depletion of pericytes around retinal vessels by tamoxifen, intra-vitreal administration of VEGF-A (1 μg), and daily intra-peritoneal administration of ABTTA (20 mg kg −1 ) for 4 days in adult DTA iΔPC mice. Analyses were performed 4 days after VEGF-A administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, dextran (70 kDa) leakage, and F4/80 + macrophage infiltration in VA-DTA iΔPC mice treated with Fc (Fc; n =6) or ABTAA (AB; n =6). Boxed regions (dotted-lines) are magnified and presented in panels in the corner. Error bars represent mean±s.d. ** P
Figure Legend Snippet: Tie2 activation lessens BRB disintegration in pericyte-free adult retina. ( a ) Diagram depicting the experiment schedule for selective depletion of pericytes around retinal vessels by tamoxifen, intra-vitreal administration of VEGF-A (1 μg), and daily intra-peritoneal administration of ABTTA (20 mg kg −1 ) for 4 days in adult DTA iΔPC mice. Analyses were performed 4 days after VEGF-A administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, dextran (70 kDa) leakage, and F4/80 + macrophage infiltration in VA-DTA iΔPC mice treated with Fc (Fc; n =6) or ABTAA (AB; n =6). Boxed regions (dotted-lines) are magnified and presented in panels in the corner. Error bars represent mean±s.d. ** P

Techniques Used: Activation Assay, Mouse Assay

2) Product Images from "Fibroblast activation protein in osteosarcoma cells promotes angiogenesis via AKT and ERK signaling pathways"

Article Title: Fibroblast activation protein in osteosarcoma cells promotes angiogenesis via AKT and ERK signaling pathways

Journal: Oncology Letters

doi: 10.3892/ol.2018.8027

FAP expression promotes VEGF-A expression in osteosarcoma cells. (A) Subsequent to the overexpression of FAP in MG63 cells, VEGF-A mRNA expression was upregulated, as determined with RT-PCR, which was (B) quantified by densitometry. (C) In addition, VEGF-A protein expression was upregulated, which was also (D) quantified. (E) Following the silencing of FAP expression in U2-OS cells, the expression of VEGF-A was inhibited at the mRNA level, as determined with RT-PCR, which was (F) quantified, and at (G) the protein level, which was (H) quantified. (I) The increase in VEGF-A subsequent to FAP overexpression and (J) the decrease in VEGF-A subsequent to FAP silencing were also confirmed by ELISA. *P
Figure Legend Snippet: FAP expression promotes VEGF-A expression in osteosarcoma cells. (A) Subsequent to the overexpression of FAP in MG63 cells, VEGF-A mRNA expression was upregulated, as determined with RT-PCR, which was (B) quantified by densitometry. (C) In addition, VEGF-A protein expression was upregulated, which was also (D) quantified. (E) Following the silencing of FAP expression in U2-OS cells, the expression of VEGF-A was inhibited at the mRNA level, as determined with RT-PCR, which was (F) quantified, and at (G) the protein level, which was (H) quantified. (I) The increase in VEGF-A subsequent to FAP overexpression and (J) the decrease in VEGF-A subsequent to FAP silencing were also confirmed by ELISA. *P

Techniques Used: Expressing, Over Expression, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

FAP and VEGF-A expression in osteosarcoma cells. FAP and VEGF-A expression were detected in MG63, U2-OS and HOS cell lines. (A) FAP and VEGF-A mRNA expression levels were determined with reverse transcription-polymerase chain reaction, which was (B) quantified by densitometry. (C) FAP and VEGF-A protein expression levels were determined with western blot analysis, which was (D) quantified by densitometry. FAP and VEGF-A in U2-OS cells was compared with the other two groups. *P
Figure Legend Snippet: FAP and VEGF-A expression in osteosarcoma cells. FAP and VEGF-A expression were detected in MG63, U2-OS and HOS cell lines. (A) FAP and VEGF-A mRNA expression levels were determined with reverse transcription-polymerase chain reaction, which was (B) quantified by densitometry. (C) FAP and VEGF-A protein expression levels were determined with western blot analysis, which was (D) quantified by densitometry. FAP and VEGF-A in U2-OS cells was compared with the other two groups. *P

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

3) Product Images from "Interaction with tumor-associated macrophages promotes PRL-3-induced invasion of colorectal cancer cells via MAPK pathway-induced EMT and NF-κB signaling-induced angiogenesis"

Article Title: Interaction with tumor-associated macrophages promotes PRL-3-induced invasion of colorectal cancer cells via MAPK pathway-induced EMT and NF-κB signaling-induced angiogenesis

Journal: Oncology Reports

doi: 10.3892/or.2019.7049

MAPK pathway regulates invasion of CRCs. (A) IL-6 and (B) IL-8 expression in supernatants from co-culture systems measured by ELISA. (C) Blocking ERK and JNK decreases the invasiveness of colorectal cancer cells. (D) A wound-healing assay showed the same trend. *P
Figure Legend Snippet: MAPK pathway regulates invasion of CRCs. (A) IL-6 and (B) IL-8 expression in supernatants from co-culture systems measured by ELISA. (C) Blocking ERK and JNK decreases the invasiveness of colorectal cancer cells. (D) A wound-healing assay showed the same trend. *P

Techniques Used: Expressing, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay, Wound Healing Assay

CCL26 increases the expression of IL-6 and IL-8 and the activation of p-JNK and p-ERK in HT29-P cells, resulting in an increase in the invasiveness of CRC cells. (A) Western blot assays and (B) densitometry analysis were performed to measure the expression of IL-6, IL-8, p-JNK, and p-ERK. (C and D) Invasion and migration were analyzed in Matrigel invasion assays. (E) IL-6 and (F) IL-8 were determined by ELISA. *P
Figure Legend Snippet: CCL26 increases the expression of IL-6 and IL-8 and the activation of p-JNK and p-ERK in HT29-P cells, resulting in an increase in the invasiveness of CRC cells. (A) Western blot assays and (B) densitometry analysis were performed to measure the expression of IL-6, IL-8, p-JNK, and p-ERK. (C and D) Invasion and migration were analyzed in Matrigel invasion assays. (E) IL-6 and (F) IL-8 were determined by ELISA. *P

Techniques Used: Expressing, Activation Assay, Western Blot, Migration, Enzyme-linked Immunosorbent Assay

4) Product Images from "Heterogeneity in VEGFR3 levels drives lymphatic vessel hyperplasia through cell-autonomous and non-cell-autonomous mechanisms"

Article Title: Heterogeneity in VEGFR3 levels drives lymphatic vessel hyperplasia through cell-autonomous and non-cell-autonomous mechanisms

Journal: Nature Communications

doi: 10.1038/s41467-018-03692-0

Vegfr3 -deleted LECs induce proliferation of neighboring VEGFR3 + cells in a cell-contact-dependent manner. a VEGF-C concentration measured by ELISA in the ear skin at indicated stages. Bars represent mean ( n = 2–8 mice, as indicated) ± s.d. b Experimental set up for co-cultures of WT and VEGFR3 − (KO) primary LECs from Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice (left) and immunofluorescence showing VEGFR3 depletion in Cre − targeted (GFP + ) LECs (right, arrows). c Left: proliferating WT cells were frequently associated with KO cells. Right: quantification of the proportion of EdU + WT cells that were in direct contact with KO cells (mean ( n = 6 biological replicates) ± s.e.m). d Assessment of cell proliferation in co-cultures of primary human LECs treated with control (siCTRL) or VEGFR3 (siVEGFR3) siRNA. Cell tracker-labeled siCTRL cells (purple) were mixed with unlabeled siVEGFR3 cells in 40:60%, or they were cultured alone. e Quantification of proliferating EdU + cells in co-cultures of siCTRL and siVEGFR3 LECs mixed in different ratios. Data are normalized to the proliferation rate (0.8 ± 0.2%) in siCTRL alone and represent mean ( n = 5 biological replicates) ± s.e.m. f Predicted and observed proportions of siCTRL LECs (all vs. EdU + ) that are in direct contact with siVEGFR3 LECs in 80:20% siCTRL:siVEGFR3 co-cultures (mean ( n = 5 biological replicates) ± s.e.m). Predicted proportion was calculated based on random distribution and n = 5 (5.2 ± 0.2, quantified from n = 172 LECs stained for VE-cadherin) neighbors. g Quantification of proliferating EdU + cells in siCTRL LECs cultured alone (−), or co-cultured without direct cell–cell contacts with siVEGFR3 cells seeded on Transwell inserts (+). Data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m. * P
Figure Legend Snippet: Vegfr3 -deleted LECs induce proliferation of neighboring VEGFR3 + cells in a cell-contact-dependent manner. a VEGF-C concentration measured by ELISA in the ear skin at indicated stages. Bars represent mean ( n = 2–8 mice, as indicated) ± s.d. b Experimental set up for co-cultures of WT and VEGFR3 − (KO) primary LECs from Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice (left) and immunofluorescence showing VEGFR3 depletion in Cre − targeted (GFP + ) LECs (right, arrows). c Left: proliferating WT cells were frequently associated with KO cells. Right: quantification of the proportion of EdU + WT cells that were in direct contact with KO cells (mean ( n = 6 biological replicates) ± s.e.m). d Assessment of cell proliferation in co-cultures of primary human LECs treated with control (siCTRL) or VEGFR3 (siVEGFR3) siRNA. Cell tracker-labeled siCTRL cells (purple) were mixed with unlabeled siVEGFR3 cells in 40:60%, or they were cultured alone. e Quantification of proliferating EdU + cells in co-cultures of siCTRL and siVEGFR3 LECs mixed in different ratios. Data are normalized to the proliferation rate (0.8 ± 0.2%) in siCTRL alone and represent mean ( n = 5 biological replicates) ± s.e.m. f Predicted and observed proportions of siCTRL LECs (all vs. EdU + ) that are in direct contact with siVEGFR3 LECs in 80:20% siCTRL:siVEGFR3 co-cultures (mean ( n = 5 biological replicates) ± s.e.m). Predicted proportion was calculated based on random distribution and n = 5 (5.2 ± 0.2, quantified from n = 172 LECs stained for VE-cadherin) neighbors. g Quantification of proliferating EdU + cells in siCTRL LECs cultured alone (−), or co-cultured without direct cell–cell contacts with siVEGFR3 cells seeded on Transwell inserts (+). Data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m. * P

Techniques Used: Concentration Assay, Enzyme-linked Immunosorbent Assay, Mouse Assay, Immunofluorescence, Labeling, Cell Culture, Staining

Global and mosaic loss of DLL4 promote LEC proliferation through inhibition of Notch signaling in neighboring cells. a , b DAPT induced effect on the proliferation of control (siCTRL) or VEGFR3 (siVEGFR3) siRNA-treated LECs. a Tile scan immunofluorescence images and ( b ) quantification of proliferating EdU + LECs after 3 h of incorporation (mean ( n = 3 biological replicates) ± s.e.m). c Western blot analysis of cell lysates from LECs treated with siCTRL or NOTCH1 siRNA (siNOTCH1) and ( d ) quantification of proliferating EdU + LECs cultured with or without VEGF-C (data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 3 biological replicates) ± s.e.m). e Western blot analysis of cell lysates from LECs treated with siCTRL or DLL4 siRNA (siDLL4) and ( f ) quantification of proliferating EdU + cells in co-cultures of siCTRL and siDLL4 LECs mixed in different ratios (data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m). g Model of cell-autonomous and non-cell-autonomous effects of VEGFR3 expression defining LEC responses during sprouting lymphangiogenesis. VEGF-C-VEGFR3 signaling positively regulates DLL4 expression leading to activation of Notch and cell cycle arrest in neighboring LECs. VEGFR3 deficiency (green cell) consequently leads to downregulation of DLL4 and inhibition of Notch in neighboring LECs, but only VEGFR3 expressing (red) cells respond by proliferation and sprouting. When VEGFR3 − cells are in excess, VEGFR3 + cells interact predominantly with LECs with low DLL4 and respond by proliferation (dashed line) * P
Figure Legend Snippet: Global and mosaic loss of DLL4 promote LEC proliferation through inhibition of Notch signaling in neighboring cells. a , b DAPT induced effect on the proliferation of control (siCTRL) or VEGFR3 (siVEGFR3) siRNA-treated LECs. a Tile scan immunofluorescence images and ( b ) quantification of proliferating EdU + LECs after 3 h of incorporation (mean ( n = 3 biological replicates) ± s.e.m). c Western blot analysis of cell lysates from LECs treated with siCTRL or NOTCH1 siRNA (siNOTCH1) and ( d ) quantification of proliferating EdU + LECs cultured with or without VEGF-C (data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 3 biological replicates) ± s.e.m). e Western blot analysis of cell lysates from LECs treated with siCTRL or DLL4 siRNA (siDLL4) and ( f ) quantification of proliferating EdU + cells in co-cultures of siCTRL and siDLL4 LECs mixed in different ratios (data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m). g Model of cell-autonomous and non-cell-autonomous effects of VEGFR3 expression defining LEC responses during sprouting lymphangiogenesis. VEGF-C-VEGFR3 signaling positively regulates DLL4 expression leading to activation of Notch and cell cycle arrest in neighboring LECs. VEGFR3 deficiency (green cell) consequently leads to downregulation of DLL4 and inhibition of Notch in neighboring LECs, but only VEGFR3 expressing (red) cells respond by proliferation and sprouting. When VEGFR3 − cells are in excess, VEGFR3 + cells interact predominantly with LECs with low DLL4 and respond by proliferation (dashed line) * P

Techniques Used: Inhibition, Immunofluorescence, Western Blot, Cell Culture, Expressing, Activation Assay

Lymphatic vascular hyperplasia in Vegfr3 -deleted ears is driven by VEGF-C signaling. a Quantification of blunt-ended vessels (lymphatic capillaries) in 5 weeks old mice treated with Tamoxifen at P2, P4, and P6. Central region excludes 560 μm area (tip) from the edge of the ear. Bars represent mean ( n = 3–6 mice, as indicated) ± s.e.m. b Whole-mount immunofluorescence of Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 ear skin showing vessel interconnections (arrows) formed of non-targeted (GFP − ) LYVE1 + (left) and VEGFR3 + (right) LECs. c Whole-mount immunofluorescence of ear skin of 3 weeks old Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 and Cre-negative littermate mice treated with Tamoxifen at P2, P4, and P6, and inhibitors of VEGF-C signaling (the VEGF-C-trap VEGFR3-Ig, the VEGFR3 inhibitor MAZ51, or the VEGFR2 blocking antibody DC101). Quantification of ( d ) vessel area and ( e ) branch points. Bars represent mean ( n = 3 (untreated groups) or n = 4 (treated groups) mice) ± s.e.m. f – h Visualization by whole-mount immunofluorescence (arrows in f ) and quantification of blunt-ended vessels in the ( g ) entire and ( h ) central region of the ear in Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice treated with the VEGFR3 kinase inhibitor MAZ51. In g , h , bars represent mean ( n = 3 (untreated) or n = 4 (MAZ51, DC101) mice) ± s.e.m. * P
Figure Legend Snippet: Lymphatic vascular hyperplasia in Vegfr3 -deleted ears is driven by VEGF-C signaling. a Quantification of blunt-ended vessels (lymphatic capillaries) in 5 weeks old mice treated with Tamoxifen at P2, P4, and P6. Central region excludes 560 μm area (tip) from the edge of the ear. Bars represent mean ( n = 3–6 mice, as indicated) ± s.e.m. b Whole-mount immunofluorescence of Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 ear skin showing vessel interconnections (arrows) formed of non-targeted (GFP − ) LYVE1 + (left) and VEGFR3 + (right) LECs. c Whole-mount immunofluorescence of ear skin of 3 weeks old Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 and Cre-negative littermate mice treated with Tamoxifen at P2, P4, and P6, and inhibitors of VEGF-C signaling (the VEGF-C-trap VEGFR3-Ig, the VEGFR3 inhibitor MAZ51, or the VEGFR2 blocking antibody DC101). Quantification of ( d ) vessel area and ( e ) branch points. Bars represent mean ( n = 3 (untreated groups) or n = 4 (treated groups) mice) ± s.e.m. f – h Visualization by whole-mount immunofluorescence (arrows in f ) and quantification of blunt-ended vessels in the ( g ) entire and ( h ) central region of the ear in Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice treated with the VEGFR3 kinase inhibitor MAZ51. In g , h , bars represent mean ( n = 3 (untreated) or n = 4 (MAZ51, DC101) mice) ± s.e.m. * P

Techniques Used: Mouse Assay, Immunofluorescence, Blocking Assay

5) Product Images from "Interactions between fibroblastic reticular cells and B cells promote mesenteric lymph node lymphangiogenesis"

Article Title: Interactions between fibroblastic reticular cells and B cells promote mesenteric lymph node lymphangiogenesis

Journal: Nature Communications

doi: 10.1038/s41467-017-00504-9

BAFF and IL-4 stimulation of B cells promotes lymphotoxin and VEGFs production. a – c mLN B cells were cultured with anti-IgM with or without IL-4±BAFF and the culture supernatant collected at a 72 h or b 18 h post stimulation for quantitation of VEGF-A and VEGF-C by ELISA. c Splenic B cells were stimulated as in ( a ) and the culture supernatant collected at 72 h for quantitation of VEGF-A and VEGF-C by ELISA. Data represent mean ± SEM, and are representative of pooled data from three independent experiments a or are representative of two independent experiments b , c . d Naive mLN B cells were cultured with or without IL-4±BAFF for 18 h then B-cell lymphotoxin expression determined by staining with LTβR-Fc. Data represent mean ± SEM and representative of three independent experiments. e mLN cryosections from mixed BMC-lacking lymphotoxin on B or T cells and infected with Hp for 21 days were stained for BAFF + ( red ) and FRCs (ER-TR7 + Laminin + ). Scale bar = 100 μm. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P
Figure Legend Snippet: BAFF and IL-4 stimulation of B cells promotes lymphotoxin and VEGFs production. a – c mLN B cells were cultured with anti-IgM with or without IL-4±BAFF and the culture supernatant collected at a 72 h or b 18 h post stimulation for quantitation of VEGF-A and VEGF-C by ELISA. c Splenic B cells were stimulated as in ( a ) and the culture supernatant collected at 72 h for quantitation of VEGF-A and VEGF-C by ELISA. Data represent mean ± SEM, and are representative of pooled data from three independent experiments a or are representative of two independent experiments b , c . d Naive mLN B cells were cultured with or without IL-4±BAFF for 18 h then B-cell lymphotoxin expression determined by staining with LTβR-Fc. Data represent mean ± SEM and representative of three independent experiments. e mLN cryosections from mixed BMC-lacking lymphotoxin on B or T cells and infected with Hp for 21 days were stained for BAFF + ( red ) and FRCs (ER-TR7 + Laminin + ). Scale bar = 100 μm. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P

Techniques Used: Cell Culture, Quantitation Assay, Enzyme-linked Immunosorbent Assay, Expressing, Staining, Infection

In vivo BAFF inhibition attenuates helminth-induced lymphangiogenesis. a C57BL/6 J wild-type mice were treated with isotype control or anti-BAFF mAb (Sandy-2) and infected with Hp . The entire chain of the mLN was collected at 12 dpi and processed for flow cytometry or histological staining. b Total weight of mLN, c total cell count, d total number of B cells, e absolute number of lymphotoxin-expressing B cells, and f total FRCs (PDPN + MadCAM1 − CD31 − ) present within the mLN as determined using flow cytometry. g mLN serial cryosections showing lymphatic organization after treatment with isotype control or anti-BAFF mAbs in naive or at 12 dpi mice ( blue ; DAPI, grays ; LYVE-1 + LECs). Scale bar = 2000 μm. Images are from a single mouse and are representative of two independent experiments each including n ≥ 2 mice/group/time point. h VEGF-A and i VEGF-C in mLN tissue homogenates as determined by ELISA. j , k total LECs (PDPN + CD-31 + ) and proliferating LECs (PDPN + CD-31 + Ki-67 + ) in the mLN of naive and 12 dpi mice treated with isotype control or anti-BAFF mAbs were determined using flow cytometry. Data represent mean ± SEM and representative of two independent experiments with n = 3mice/group/time-point. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P
Figure Legend Snippet: In vivo BAFF inhibition attenuates helminth-induced lymphangiogenesis. a C57BL/6 J wild-type mice were treated with isotype control or anti-BAFF mAb (Sandy-2) and infected with Hp . The entire chain of the mLN was collected at 12 dpi and processed for flow cytometry or histological staining. b Total weight of mLN, c total cell count, d total number of B cells, e absolute number of lymphotoxin-expressing B cells, and f total FRCs (PDPN + MadCAM1 − CD31 − ) present within the mLN as determined using flow cytometry. g mLN serial cryosections showing lymphatic organization after treatment with isotype control or anti-BAFF mAbs in naive or at 12 dpi mice ( blue ; DAPI, grays ; LYVE-1 + LECs). Scale bar = 2000 μm. Images are from a single mouse and are representative of two independent experiments each including n ≥ 2 mice/group/time point. h VEGF-A and i VEGF-C in mLN tissue homogenates as determined by ELISA. j , k total LECs (PDPN + CD-31 + ) and proliferating LECs (PDPN + CD-31 + Ki-67 + ) in the mLN of naive and 12 dpi mice treated with isotype control or anti-BAFF mAbs were determined using flow cytometry. Data represent mean ± SEM and representative of two independent experiments with n = 3mice/group/time-point. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P

Techniques Used: In Vivo, Inhibition, Mouse Assay, Infection, Flow Cytometry, Cytometry, Staining, Cell Counting, Expressing, Enzyme-linked Immunosorbent Assay

6) Product Images from "Metformin prevents hormonal and metabolic disturbances and 1,2-dimethylhydrazine-induced colon carcinogenesis in non-diabetic rats"

Article Title: Metformin prevents hormonal and metabolic disturbances and 1,2-dimethylhydrazine-induced colon carcinogenesis in non-diabetic rats

Journal: Cancer Biology & Medicine

doi: 10.20892/j.issn.2095-3941.2016.0088

Effect of 1,2-dimethylhydrazine (DMH) and metformin on hormonal and metabolic parameters in the serum male Wistar rats. (A) glucose. (B) Insulin. (C) IGF-1. (D) Total cholesterol. (E) Triglycerides. (F) Cu, Zn-superoxide dismutase. (G) Catalase. (H) Malonic dialdehyde. (I) Hemoglobin. (J) Glycated hemoglobin. (K) Alaninaminetransferase. (L) Aspartataminetransferase. (M) VEGF. Data presented as mean±SEM, n =6–15 per group. P ≤0.05. a: DMH vs . control; b: DMH+MF vs . DMH. Rats bearing DMH-induced colon adenocarcinomas have elevated serum level of glucose, insulin, IGF-1, total cholesterol, triglycerides, catalase, malonic dialdehyde, glycated hemoglobin, AST, ALT and decreased level of hemoglobin. Treatment with MF in both doses normalized majority of these changes in DMH-treated group of rats, whereas failed to modify them in rats not treated with DMH. Only exception was decreased level of triglycerides in MF-treated rats (Figure 1E, P
Figure Legend Snippet: Effect of 1,2-dimethylhydrazine (DMH) and metformin on hormonal and metabolic parameters in the serum male Wistar rats. (A) glucose. (B) Insulin. (C) IGF-1. (D) Total cholesterol. (E) Triglycerides. (F) Cu, Zn-superoxide dismutase. (G) Catalase. (H) Malonic dialdehyde. (I) Hemoglobin. (J) Glycated hemoglobin. (K) Alaninaminetransferase. (L) Aspartataminetransferase. (M) VEGF. Data presented as mean±SEM, n =6–15 per group. P ≤0.05. a: DMH vs . control; b: DMH+MF vs . DMH. Rats bearing DMH-induced colon adenocarcinomas have elevated serum level of glucose, insulin, IGF-1, total cholesterol, triglycerides, catalase, malonic dialdehyde, glycated hemoglobin, AST, ALT and decreased level of hemoglobin. Treatment with MF in both doses normalized majority of these changes in DMH-treated group of rats, whereas failed to modify them in rats not treated with DMH. Only exception was decreased level of triglycerides in MF-treated rats (Figure 1E, P

Techniques Used: AST Assay

7) Product Images from "Ethyl-p-methoxycinnamate isolated from kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-?, and angiogenesis by blocking endothelial functions"

Article Title: Ethyl-p-methoxycinnamate isolated from kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-?, and angiogenesis by blocking endothelial functions

Journal: Clinics

doi: 10.6061/clinics/2014(02)10

Enzyme-linked immunosorbant assay to measure TNF-α and IL-1
Figure Legend Snippet: Enzyme-linked immunosorbant assay to measure TNF-α and IL-1

Techniques Used:

A ) The mean weight of cotton pellets dissected from rats treated with ethyl- p -methoxycinnamate (EPMC) at doses of 200, 400, and 800 mg/kg. B ) Percent increase in the time taken by rats to flick their tail away from heat, a measure of the analgesic effect of EPMC, at doses of 200, 400, and 800 mg/kg. C ) Blood concentration of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) in rats treated with EPMC at doses of 200, 400, and 800 mg/kg on the 8 th day of the cotton pellet granuloma assay. D ) Percent inhibition of IL-1, TNF-α, and nitric oxide (NO) synthesis by EPMC in U937 cells at concentrations of 25, 50, 100, and 200 µg/ml. The cell culture supernatants were collected after 48 hours and subjected to an enzyme-linked immunosorbant assay (ELISA) for the assessment of IL-1, TNF-α, and NO. All values are represented as the mean ± SEM ( n = 6), and * , ** , and *** indicate significant differences of p
Figure Legend Snippet: A ) The mean weight of cotton pellets dissected from rats treated with ethyl- p -methoxycinnamate (EPMC) at doses of 200, 400, and 800 mg/kg. B ) Percent increase in the time taken by rats to flick their tail away from heat, a measure of the analgesic effect of EPMC, at doses of 200, 400, and 800 mg/kg. C ) Blood concentration of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) in rats treated with EPMC at doses of 200, 400, and 800 mg/kg on the 8 th day of the cotton pellet granuloma assay. D ) Percent inhibition of IL-1, TNF-α, and nitric oxide (NO) synthesis by EPMC in U937 cells at concentrations of 25, 50, 100, and 200 µg/ml. The cell culture supernatants were collected after 48 hours and subjected to an enzyme-linked immunosorbant assay (ELISA) for the assessment of IL-1, TNF-α, and NO. All values are represented as the mean ± SEM ( n = 6), and * , ** , and *** indicate significant differences of p

Techniques Used: Concentration Assay, Inhibition, Cell Culture, Enzyme-linked Immunosorbent Assay

8) Product Images from "The Effect of Age on the Regenerative Potential of Human Eyelid Adipose-Derived Stem Cells"

Article Title: The Effect of Age on the Regenerative Potential of Human Eyelid Adipose-Derived Stem Cells

Journal: Stem Cells International

doi: 10.1155/2018/5654917

ELISA analysis of secreted proteins in HEASCs-CM. The graph showed that the expression of TGF- β 1 and cFn was decreased with advanced age; however, the expression of VEGF demonstrated an age-related increase in donor age groups. Data was represented as mean ± SD, ∗ P
Figure Legend Snippet: ELISA analysis of secreted proteins in HEASCs-CM. The graph showed that the expression of TGF- β 1 and cFn was decreased with advanced age; however, the expression of VEGF demonstrated an age-related increase in donor age groups. Data was represented as mean ± SD, ∗ P

Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing

9) Product Images from "Therapeutic Lymphangiogenesis With Implantation of Adipose-Derived Regenerative Cells"

Article Title: Therapeutic Lymphangiogenesis With Implantation of Adipose-Derived Regenerative Cells

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

doi: 10.1161/JAHA.112.000877

Implantation of ADRCs and lymphangiogenic cytokines in vivo. A, The abundance of VEGF‐C and HGF mRNA in the ADRC group was significantly greater than that of the control group by real‐time reverse transcriptase–polymerase chain reaction (VEGF‐C: 6.7‐fold, n=5, P
Figure Legend Snippet: Implantation of ADRCs and lymphangiogenic cytokines in vivo. A, The abundance of VEGF‐C and HGF mRNA in the ADRC group was significantly greater than that of the control group by real‐time reverse transcriptase–polymerase chain reaction (VEGF‐C: 6.7‐fold, n=5, P

Techniques Used: In Vivo, Polymerase Chain Reaction

The ability of ADRCs for lymphangiogenesis in vitro (functional assay and differentiation assay). A, Enzyme‐linked immunosorbent assay (ELISA) revealed that the concentration of VEGF‐C was upregulated in ADRC‐CM compared to control. B, rhVEGF‐C induced LEC migration in a dose‐dependent manner, and ADRC‐CM also induced LEC migration. HPF indicates high‐powered field. * P
Figure Legend Snippet: The ability of ADRCs for lymphangiogenesis in vitro (functional assay and differentiation assay). A, Enzyme‐linked immunosorbent assay (ELISA) revealed that the concentration of VEGF‐C was upregulated in ADRC‐CM compared to control. B, rhVEGF‐C induced LEC migration in a dose‐dependent manner, and ADRC‐CM also induced LEC migration. HPF indicates high‐powered field. * P

Techniques Used: In Vitro, Functional Assay, Differentiation Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Migration

Newly established mouse model of tail lymphedema. A, Circumferential annulus of skin (2 mm wide) located 10 mm distal to the tail base, excluding a 4‐mm 2 dermal flap at the ventral side, was removed from the tail. Good blood perfusion was maintained at distal site. B, Tail lymphedema was induced within a few days after the procedure, and the tail diameter peaked at around postoperative day 7. Lymphedema continued in the PBS group to at least 28 days. C, Representative photomicrographs of histological sections in lymphedematous tail tissues distal to the incision site. The space of subcutaneous tissues was edematous and dilated in the PBS group, but the space was almost normal in the ADRC implantation group (double‐headed arrow). Scale bar=2 mm. D, As a positive control, a marked reduction of the edema was observed after day 11 in VEGF‐C protein–administered group compared to PBS group ( P
Figure Legend Snippet: Newly established mouse model of tail lymphedema. A, Circumferential annulus of skin (2 mm wide) located 10 mm distal to the tail base, excluding a 4‐mm 2 dermal flap at the ventral side, was removed from the tail. Good blood perfusion was maintained at distal site. B, Tail lymphedema was induced within a few days after the procedure, and the tail diameter peaked at around postoperative day 7. Lymphedema continued in the PBS group to at least 28 days. C, Representative photomicrographs of histological sections in lymphedematous tail tissues distal to the incision site. The space of subcutaneous tissues was edematous and dilated in the PBS group, but the space was almost normal in the ADRC implantation group (double‐headed arrow). Scale bar=2 mm. D, As a positive control, a marked reduction of the edema was observed after day 11 in VEGF‐C protein–administered group compared to PBS group ( P

Techniques Used: Positive Control

Possible mechanisms of lymphangiogenesis mediated by ADRC implantation. We propose 2 main mechanisms: First, implanted ADRCs release cytokines, including VEGF‐C, that might stimulate migration and proliferation of residual LECs and eventual lymphangiogenesis. Second, cytokines released from ADRCs could augment mobilization and/or recruitment of bone marrow–derived M2 macrophages serving as lymphatic endothelial progenitors. There is little evidence that implanted ADRCs directly transdifferentiate into mature LECs.
Figure Legend Snippet: Possible mechanisms of lymphangiogenesis mediated by ADRC implantation. We propose 2 main mechanisms: First, implanted ADRCs release cytokines, including VEGF‐C, that might stimulate migration and proliferation of residual LECs and eventual lymphangiogenesis. Second, cytokines released from ADRCs could augment mobilization and/or recruitment of bone marrow–derived M2 macrophages serving as lymphatic endothelial progenitors. There is little evidence that implanted ADRCs directly transdifferentiate into mature LECs.

Techniques Used: Migration, Derivative Assay

10) Product Images from "The Evaluation of the Relationship between sTREM-1, VEGF-B, and VEGF Gene Expression Levels with Disease Activity of Behçet's Patients"

Article Title: The Evaluation of the Relationship between sTREM-1, VEGF-B, and VEGF Gene Expression Levels with Disease Activity of Behçet's Patients

Journal: Disease Markers

doi: 10.1155/2018/2649392

The sTREM-1, VEGF-B, and VEGF gene expression levels of Behçet's disease patients and healthy controls.
Figure Legend Snippet: The sTREM-1, VEGF-B, and VEGF gene expression levels of Behçet's disease patients and healthy controls.

Techniques Used: Expressing

11) Product Images from "Distinct Characteristics of Circulating Vascular Endothelial Growth Factor-A and C Levels in Human Subjects"

Article Title: Distinct Characteristics of Circulating Vascular Endothelial Growth Factor-A and C Levels in Human Subjects

Journal: PLoS ONE

doi: 10.1371/journal.pone.0029351

The correlation of circulating vascular endothelial growth factor-A (VEGF-A) or C (VEGF-C) levels with their independent determinants. A. The correlation between circulating VEGF-A levels and the body mass index. B. The correlation between those of VEGF-C and those of triglycerides. C. The correlation between those of VEGF-C and those of non-high-density-lipoprotein cholesterol (nonHDL-C).
Figure Legend Snippet: The correlation of circulating vascular endothelial growth factor-A (VEGF-A) or C (VEGF-C) levels with their independent determinants. A. The correlation between circulating VEGF-A levels and the body mass index. B. The correlation between those of VEGF-C and those of triglycerides. C. The correlation between those of VEGF-C and those of non-high-density-lipoprotein cholesterol (nonHDL-C).

Techniques Used:

Serum and expression levels in atheromatous plaque of VEGF-A and VEGF-C in apoE-deficient mice. A. Quantification of the lesion size in the proximal aortas of apolipoprotein E (apoE)-deficient mice fed normal chow (NC, n = 3) or a high-fat-diet (HFD, n = 3). The ratio of the atherosclerotic area to the total area was significantly greater in HFD than NC mice. B. Quantification of the expression of vascular endothelial growth factor-A (VEGF-A) and vascular endothelial growth factor-C (VEGF-C) in NC and HFD mice. The expression of VEGF-C, but not VEGF-A, was significantly intensified by feeding HFD compared to NC. C–F. Representative microscopic views (x400) of the expression of VEGF-A in the aortic sinus of apoE-deficient mice fed NC (C) or a HFD (D), and those of VEGF-C in NC (E) or HFD (F) mice. The red arrows indicate VEGF-A- or VEGF-C-positive cells. G and H. Serum levels of VEGF-A (G) and VEGF-C (H) in apoE-deficient mice fed a HFD or NC for 16 weeks. The data are means ± SD.
Figure Legend Snippet: Serum and expression levels in atheromatous plaque of VEGF-A and VEGF-C in apoE-deficient mice. A. Quantification of the lesion size in the proximal aortas of apolipoprotein E (apoE)-deficient mice fed normal chow (NC, n = 3) or a high-fat-diet (HFD, n = 3). The ratio of the atherosclerotic area to the total area was significantly greater in HFD than NC mice. B. Quantification of the expression of vascular endothelial growth factor-A (VEGF-A) and vascular endothelial growth factor-C (VEGF-C) in NC and HFD mice. The expression of VEGF-C, but not VEGF-A, was significantly intensified by feeding HFD compared to NC. C–F. Representative microscopic views (x400) of the expression of VEGF-A in the aortic sinus of apoE-deficient mice fed NC (C) or a HFD (D), and those of VEGF-C in NC (E) or HFD (F) mice. The red arrows indicate VEGF-A- or VEGF-C-positive cells. G and H. Serum levels of VEGF-A (G) and VEGF-C (H) in apoE-deficient mice fed a HFD or NC for 16 weeks. The data are means ± SD.

Techniques Used: Expressing, Mouse Assay

12) Product Images from "miR-129-5p targets Wnt5a to block PKC/ERK/NF-κB and JNK pathways in glioblastoma"

Article Title: miR-129-5p targets Wnt5a to block PKC/ERK/NF-κB and JNK pathways in glioblastoma

Journal: Cell Death & Disease

doi: 10.1038/s41419-018-0343-1

miR-129-5p overexpression inhibits cell proliferation and angiogenesis in GBM by targeting Wnt5a . a N3/miR-129-5p, K3/miR-129-5p and U251/miR-129-5p cells were transfected with the Wnt5a-plasmid vector, followed by western blot analysis of Wnt5a transcripts (three replicates per group, three independent experiments per group). GAPDH served as the loading control. b Colony formation ability of the miR-NC- or miR-129-5p-transfected N3 or U251 cells without transfection or transfected with the pcDNA3.1-Wnt5a plasmid (Wnt5a) (six replicates per group, three independent experiments per group). Scale bar, 2.5 mm. c Overexpression of miR-129-5p arrested cell proliferation; however, this was rescued upon coexpression of exogenous Wnt5a in N3 and U251 cells (six replicates per group, three independent experiments per group). d EdU analysis of miR-NC, miR-129-5p or miR-129-5p plus Wnt5a-transfected N3 and U251 cells (six replicates per group, three independent experiments per group). Scale bar, 100 μm. e Cell-cycle assay of N3 and U251 GBM cells 3 days after transfection with miR-NC, miR-129-5p or miR-129-5p plus Wnt5a (six replicates per group, three independent experiments per group). f Western blot analysis indicated the regulation of the cell-cycle-regulatory proteins cyclin E1 and cyclin D1 in miR-NC, miR-129-5p or miR-129-5p mimic plus Wnt5a-transfected N3 or U251 cells. GAPDH was used as the loading control (three replicates per group, three independent experiments per group). g Representative images and quantification of HBMVECs cultured on Matrigel-coated 96-well plates using conditioned medium from the indicated cells (six replicates per group, three independent experiments per group). Scale bar, 200 μm. Data are expressed as the mean ± s.e.m. h – i Quantitative determination of Wnt5a and VEGF levels in supernatants from N3 or U251 cells with indicated treatment by ELISA. (six replicates per group, three independent experiments per group). Data are expressed as the mean ± s.e.m
Figure Legend Snippet: miR-129-5p overexpression inhibits cell proliferation and angiogenesis in GBM by targeting Wnt5a . a N3/miR-129-5p, K3/miR-129-5p and U251/miR-129-5p cells were transfected with the Wnt5a-plasmid vector, followed by western blot analysis of Wnt5a transcripts (three replicates per group, three independent experiments per group). GAPDH served as the loading control. b Colony formation ability of the miR-NC- or miR-129-5p-transfected N3 or U251 cells without transfection or transfected with the pcDNA3.1-Wnt5a plasmid (Wnt5a) (six replicates per group, three independent experiments per group). Scale bar, 2.5 mm. c Overexpression of miR-129-5p arrested cell proliferation; however, this was rescued upon coexpression of exogenous Wnt5a in N3 and U251 cells (six replicates per group, three independent experiments per group). d EdU analysis of miR-NC, miR-129-5p or miR-129-5p plus Wnt5a-transfected N3 and U251 cells (six replicates per group, three independent experiments per group). Scale bar, 100 μm. e Cell-cycle assay of N3 and U251 GBM cells 3 days after transfection with miR-NC, miR-129-5p or miR-129-5p plus Wnt5a (six replicates per group, three independent experiments per group). f Western blot analysis indicated the regulation of the cell-cycle-regulatory proteins cyclin E1 and cyclin D1 in miR-NC, miR-129-5p or miR-129-5p mimic plus Wnt5a-transfected N3 or U251 cells. GAPDH was used as the loading control (three replicates per group, three independent experiments per group). g Representative images and quantification of HBMVECs cultured on Matrigel-coated 96-well plates using conditioned medium from the indicated cells (six replicates per group, three independent experiments per group). Scale bar, 200 μm. Data are expressed as the mean ± s.e.m. h – i Quantitative determination of Wnt5a and VEGF levels in supernatants from N3 or U251 cells with indicated treatment by ELISA. (six replicates per group, three independent experiments per group). Data are expressed as the mean ± s.e.m

Techniques Used: Over Expression, Transfection, Plasmid Preparation, Western Blot, Cell Cycle Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

13) Product Images from "Unexpected contribution of lymphatic vessels to promotion of distant metastatic tumor spread"

Article Title: Unexpected contribution of lymphatic vessels to promotion of distant metastatic tumor spread

Journal: Science Advances

doi: 10.1126/sciadv.aat4758

Characterization of CCSP-rtTA × tet-O–VEGF-C mice after doxycycline treatment for 2 weeks. ( A ) VEGF-C mRNA expression in the lung (normalized to RPLP0). The average expression level in WT × tet-O–VEGF-C mice was set to 1 ( n = 6). ( B ) VEGF-C protein levels in the lung detected by enzyme-linked immunosorbent assay (ELISA) ( n = 6). ( C ) Quantification of the VEGFR3 + LV area ( n = 4). ( D ) Representative images of VEGFR3 staining in the lung. Scale bars, 1 mm. ( E ) Representative high-magnification images of different regions in the lung stained for VEGFR3. Scale bars, 200 μm. PA, pulmonary artery; PV, pulmonary vein. ( F ) Representative images of MECA-32 staining in the lung. Scale bars, 50 μm. ( G ) Quantification of MECA-32 + blood vessel area in the lung ( n = 5).
Figure Legend Snippet: Characterization of CCSP-rtTA × tet-O–VEGF-C mice after doxycycline treatment for 2 weeks. ( A ) VEGF-C mRNA expression in the lung (normalized to RPLP0). The average expression level in WT × tet-O–VEGF-C mice was set to 1 ( n = 6). ( B ) VEGF-C protein levels in the lung detected by enzyme-linked immunosorbent assay (ELISA) ( n = 6). ( C ) Quantification of the VEGFR3 + LV area ( n = 4). ( D ) Representative images of VEGFR3 staining in the lung. Scale bars, 1 mm. ( E ) Representative high-magnification images of different regions in the lung stained for VEGFR3. Scale bars, 200 μm. PA, pulmonary artery; PV, pulmonary vein. ( F ) Representative images of MECA-32 staining in the lung. Scale bars, 50 μm. ( G ) Quantification of MECA-32 + blood vessel area in the lung ( n = 5).

Techniques Used: Mouse Assay, Expressing, Enzyme-linked Immunosorbent Assay, Staining

14) Product Images from "Bone mesenchymal stem cells ameliorate ischemia/reperfusion-induced damage in renal epithelial cells via microRNA-223"

Article Title: Bone mesenchymal stem cells ameliorate ischemia/reperfusion-induced damage in renal epithelial cells via microRNA-223

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-017-0599-x

Knockdown of miR-223 blocked the protective effect of mesenchymal stem cells ( MSCs ) on renal tubular epithelial cells ( RTECs ). After the kidney ischemia/reperfusion ( IR ) KM/NIH mouse models were established, the mice were abdominally intravenously injected with MSCs, miR-223 inhibitor ( miR-223 in ) transfected MSCs or hypoxia-pretreated MSCs ( htMSCs ). Kidney tissues were harvested at 24 h after injection. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in blood samples were measured with ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ) and cysteine protease protein-3 ( caspase-3 ) in kidney tissue was detected by qRT-PCR. c Representative images from Western blot assays for apoptosis-related indicators. a P
Figure Legend Snippet: Knockdown of miR-223 blocked the protective effect of mesenchymal stem cells ( MSCs ) on renal tubular epithelial cells ( RTECs ). After the kidney ischemia/reperfusion ( IR ) KM/NIH mouse models were established, the mice were abdominally intravenously injected with MSCs, miR-223 inhibitor ( miR-223 in ) transfected MSCs or hypoxia-pretreated MSCs ( htMSCs ). Kidney tissues were harvested at 24 h after injection. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in blood samples were measured with ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ) and cysteine protease protein-3 ( caspase-3 ) in kidney tissue was detected by qRT-PCR. c Representative images from Western blot assays for apoptosis-related indicators. a P

Techniques Used: Mouse Assay, Injection, Transfection, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot

Expression of growth factors, chemokines, and antiapoptosis factors was suppressed by coculture with mesenchymal stem cells ( MSCs ) when miR-223 was inhibited; however, expression of proapoptosis factors was enhanced. All renal tubular epithelial cells ( RTECs ) were treated with hypoxia/reoxygenation ( HR ) stimulation. The MSCs had been transfected in advance with a miR-223 inhibitor ( miR-223 in ). The RTECs and MSCs were then cocultured in a double-chamber. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in coculture supernatants were measured with ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ), and cysteine protease protein-3 ( caspase-3 ) in RTECs was detected by qRT-PCR. c Representative images of Western blot assays for apoptosis-related indicators. a P
Figure Legend Snippet: Expression of growth factors, chemokines, and antiapoptosis factors was suppressed by coculture with mesenchymal stem cells ( MSCs ) when miR-223 was inhibited; however, expression of proapoptosis factors was enhanced. All renal tubular epithelial cells ( RTECs ) were treated with hypoxia/reoxygenation ( HR ) stimulation. The MSCs had been transfected in advance with a miR-223 inhibitor ( miR-223 in ). The RTECs and MSCs were then cocultured in a double-chamber. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in coculture supernatants were measured with ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ), and cysteine protease protein-3 ( caspase-3 ) in RTECs was detected by qRT-PCR. c Representative images of Western blot assays for apoptosis-related indicators. a P

Techniques Used: Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Expression of growth factors, chemokines, and antiapoptosis factors was enhanced by coculture with mesenchymal stem cells ( MSCs ), while expression of proapoptosis factors was suppressed. Renal tubular epithelial cells ( RTECs ) were treated with hypoxia/reoxygenation ( HR ) stimulation and then cocultured with MSCs or hypoxia-pretreated MSCs ( htMSCs ) in a double-chamber. Cells were harvested at 48 h. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in coculture supernatants were measured by ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ), and cysteine protease protein-3 ( caspase-3 ) in RTECs was detected by qRT-PCR. c Representative images of Western blot assays for apoptosis-related indicators. a P
Figure Legend Snippet: Expression of growth factors, chemokines, and antiapoptosis factors was enhanced by coculture with mesenchymal stem cells ( MSCs ), while expression of proapoptosis factors was suppressed. Renal tubular epithelial cells ( RTECs ) were treated with hypoxia/reoxygenation ( HR ) stimulation and then cocultured with MSCs or hypoxia-pretreated MSCs ( htMSCs ) in a double-chamber. Cells were harvested at 48 h. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in coculture supernatants were measured by ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ), and cysteine protease protein-3 ( caspase-3 ) in RTECs was detected by qRT-PCR. c Representative images of Western blot assays for apoptosis-related indicators. a P

Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

15) Product Images from "Metformin prevents hormonal and metabolic disturbances and 1,2-dimethylhydrazine-induced colon carcinogenesis in non-diabetic rats"

Article Title: Metformin prevents hormonal and metabolic disturbances and 1,2-dimethylhydrazine-induced colon carcinogenesis in non-diabetic rats

Journal: Cancer Biology & Medicine

doi: 10.20892/j.issn.2095-3941.2016.0088

Effect of 1,2-dimethylhydrazine (DMH) and metformin on hormonal and metabolic parameters in the serum male Wistar rats. (A) glucose. (B) Insulin. (C) IGF-1. (D) Total cholesterol. (E) Triglycerides. (F) Cu, Zn-superoxide dismutase. (G) Catalase. (H) Malonic dialdehyde. (I) Hemoglobin. (J) Glycated hemoglobin. (K) Alaninaminetransferase. (L) Aspartataminetransferase. (M) VEGF. Data presented as mean±SEM, n =6–15 per group. P ≤0.05. a: DMH vs . control; b: DMH+MF vs . DMH. Rats bearing DMH-induced colon adenocarcinomas have elevated serum level of glucose, insulin, IGF-1, total cholesterol, triglycerides, catalase, malonic dialdehyde, glycated hemoglobin, AST, ALT and decreased level of hemoglobin. Treatment with MF in both doses normalized majority of these changes in DMH-treated group of rats, whereas failed to modify them in rats not treated with DMH. Only exception was decreased level of triglycerides in MF-treated rats (Figure 1E, P
Figure Legend Snippet: Effect of 1,2-dimethylhydrazine (DMH) and metformin on hormonal and metabolic parameters in the serum male Wistar rats. (A) glucose. (B) Insulin. (C) IGF-1. (D) Total cholesterol. (E) Triglycerides. (F) Cu, Zn-superoxide dismutase. (G) Catalase. (H) Malonic dialdehyde. (I) Hemoglobin. (J) Glycated hemoglobin. (K) Alaninaminetransferase. (L) Aspartataminetransferase. (M) VEGF. Data presented as mean±SEM, n =6–15 per group. P ≤0.05. a: DMH vs . control; b: DMH+MF vs . DMH. Rats bearing DMH-induced colon adenocarcinomas have elevated serum level of glucose, insulin, IGF-1, total cholesterol, triglycerides, catalase, malonic dialdehyde, glycated hemoglobin, AST, ALT and decreased level of hemoglobin. Treatment with MF in both doses normalized majority of these changes in DMH-treated group of rats, whereas failed to modify them in rats not treated with DMH. Only exception was decreased level of triglycerides in MF-treated rats (Figure 1E, P

Techniques Used: AST Assay

16) Product Images from "The Effect of Age on the Regenerative Potential of Human Eyelid Adipose-Derived Stem Cells"

Article Title: The Effect of Age on the Regenerative Potential of Human Eyelid Adipose-Derived Stem Cells

Journal: Stem Cells International

doi: 10.1155/2018/5654917

ELISA analysis of secreted proteins in HEASCs-CM. The graph showed that the expression of TGF- β 1 and cFn was decreased with advanced age; however, the expression of VEGF demonstrated an age-related increase in donor age groups. Data was represented as mean ± SD, ∗ P
Figure Legend Snippet: ELISA analysis of secreted proteins in HEASCs-CM. The graph showed that the expression of TGF- β 1 and cFn was decreased with advanced age; however, the expression of VEGF demonstrated an age-related increase in donor age groups. Data was represented as mean ± SD, ∗ P

Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing

17) Product Images from "VEGF-C promotes the development of lymphatics in bone and bone loss"

Article Title: VEGF-C promotes the development of lymphatics in bone and bone loss

Journal: eLife

doi: 10.7554/eLife.34323

Bone lymphatics in Osx-tTA;TetO-Vegfc mice disappear following the withdrawal of VEGF-C. ( A ) Schematic showing when mice received normal water and doxycycline water. One cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d). A second cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P38 (On 35d/Off 3d). A third cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P42 (On 35d/Off 7d). ( B ) Graph showing the relative VEGF-C mRNA levels in tibias from mice. ( C ) Schematic showing when mice received normal water and doxycycline water. Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d) or normal water from E18.5 to P35 and then doxycycline water for 3 (On 35d/Off 3d), 7 (On 35d/Off 7d), 28 (On 35d/Off 28d), or 56 days (On 35d/Off 56d). ( D–I ) Representative images of ribs stained with an anti-podoplanin antibody. The dashed lines separate the bone from the periosseous muscle. ( J ) Graph showing lymphatic vessel index values for ribs in Osx-tTA mice (0 ± 0.0; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (152.5 ± 29.56; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (62.25 ± 51.7; n = 4), 7 (24.08 ± 21.26; n = 4), 28 (0 ± 0.0; n = 5) or 56 (0 ± 0.0; n = 3) days. ( K ) Graph showing lymphatic vessel index values for periosseous muscle in Osx-tTA mice (3.61 ± 1.974; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (92.45 ± 34.63; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (99.29 ± 23.37; n = 4), 7 (74.84 ± 18.98; n = 4), 28 (92.67 ± 24.2; n = 5) or 56 (72.17 ± 14.05; n = 3) days. (**p
Figure Legend Snippet: Bone lymphatics in Osx-tTA;TetO-Vegfc mice disappear following the withdrawal of VEGF-C. ( A ) Schematic showing when mice received normal water and doxycycline water. One cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d). A second cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P38 (On 35d/Off 3d). A third cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P42 (On 35d/Off 7d). ( B ) Graph showing the relative VEGF-C mRNA levels in tibias from mice. ( C ) Schematic showing when mice received normal water and doxycycline water. Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d) or normal water from E18.5 to P35 and then doxycycline water for 3 (On 35d/Off 3d), 7 (On 35d/Off 7d), 28 (On 35d/Off 28d), or 56 days (On 35d/Off 56d). ( D–I ) Representative images of ribs stained with an anti-podoplanin antibody. The dashed lines separate the bone from the periosseous muscle. ( J ) Graph showing lymphatic vessel index values for ribs in Osx-tTA mice (0 ± 0.0; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (152.5 ± 29.56; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (62.25 ± 51.7; n = 4), 7 (24.08 ± 21.26; n = 4), 28 (0 ± 0.0; n = 5) or 56 (0 ± 0.0; n = 3) days. ( K ) Graph showing lymphatic vessel index values for periosseous muscle in Osx-tTA mice (3.61 ± 1.974; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (92.45 ± 34.63; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (99.29 ± 23.37; n = 4), 7 (74.84 ± 18.98; n = 4), 28 (92.67 ± 24.2; n = 5) or 56 (72.17 ± 14.05; n = 3) days. (**p

Techniques Used: Mouse Assay, Staining

Lymphatics in soft tissues in Osx-tTA;TetO-Vegfc mice appear normal. ( A ) Representative pictures of tissues collected from P35 Osx-tTA and Osx-tTA;TetO-Vegfc mice. Lymphatics in the lung, liver, kidney, and pancreas appeared normal in Osx-tTA;TetO-Vegfc mice. ( B ) Graph showing the circulating level of VEGF-C in P35 Osx-tTA (2.04 ± 1.42, n = 8) and Osx-tTA;TetO-Vegfc (2.387 ± 0.94, n = 7) mice.
Figure Legend Snippet: Lymphatics in soft tissues in Osx-tTA;TetO-Vegfc mice appear normal. ( A ) Representative pictures of tissues collected from P35 Osx-tTA and Osx-tTA;TetO-Vegfc mice. Lymphatics in the lung, liver, kidney, and pancreas appeared normal in Osx-tTA;TetO-Vegfc mice. ( B ) Graph showing the circulating level of VEGF-C in P35 Osx-tTA (2.04 ± 1.42, n = 8) and Osx-tTA;TetO-Vegfc (2.387 ± 0.94, n = 7) mice.

Techniques Used: Mouse Assay

Expression of VEGF-C during embryonic development causes embryonic lethality. ( A ) Schematic showing when mice received normal water. ( B–C ) Representative images of E12.5, E14.5 and E16.5 control embryos. ( E–G ) Representative images of E12.5, E14.5 and E16.5 Osx-tTA;TetO-Vegfc embryos. ( H,I ) Transverse sections of E14.5 embryos stained with hematoxylin and eosin. The jugular lymph sac (asterisk) in the control embryo appears normal. In contrast, the jugular lymph sac (asterisk) in the Osx-tTA;TetO-Vegfc embryo is enlarged.
Figure Legend Snippet: Expression of VEGF-C during embryonic development causes embryonic lethality. ( A ) Schematic showing when mice received normal water. ( B–C ) Representative images of E12.5, E14.5 and E16.5 control embryos. ( E–G ) Representative images of E12.5, E14.5 and E16.5 Osx-tTA;TetO-Vegfc embryos. ( H,I ) Transverse sections of E14.5 embryos stained with hematoxylin and eosin. The jugular lymph sac (asterisk) in the control embryo appears normal. In contrast, the jugular lymph sac (asterisk) in the Osx-tTA;TetO-Vegfc embryo is enlarged.

Techniques Used: Expressing, Mouse Assay, Staining

18) Product Images from "Adenosine A2a receptor promotes lymphangiogenesis and lymph node metastasis"

Article Title: Adenosine A2a receptor promotes lymphangiogenesis and lymph node metastasis

Journal: Oncoimmunology

doi: 10.1080/2162402X.2019.1601481

VEGF-C levels are reduced in A2a-deficient sentinel lymph nodes. WT and A2a-deficient mice were injected with 2.5 × 10 5 B16-CD73+ melanoma cells in the foot pad and two weeks later tumor-draining lymph nodes and contralateral lymph nodes were collected. Primary tumors and lymph node were homogenized, centrifuged and supernatants were assayed for VEGF levels by ELISA. (a) Fold-increase in VEGF-A, C and D in tumor-draining lymph nodes (TdLN) over matched contraleral lymph nodes (cLN). (b) Levels of VEGF-A, C and D in tumors. Means ± standard errors are shown. * p
Figure Legend Snippet: VEGF-C levels are reduced in A2a-deficient sentinel lymph nodes. WT and A2a-deficient mice were injected with 2.5 × 10 5 B16-CD73+ melanoma cells in the foot pad and two weeks later tumor-draining lymph nodes and contralateral lymph nodes were collected. Primary tumors and lymph node were homogenized, centrifuged and supernatants were assayed for VEGF levels by ELISA. (a) Fold-increase in VEGF-A, C and D in tumor-draining lymph nodes (TdLN) over matched contraleral lymph nodes (cLN). (b) Levels of VEGF-A, C and D in tumors. Means ± standard errors are shown. * p

Techniques Used: Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay

The number of VEGF-C secreting B cells is reduced in A2a-deficient sentinel lymph nodes. WT and A2a-deficient mice were injected with 2.5 × 10 5 B16-CD73+ melanoma cells in the foot pad. (a) Two weeks later, tumor-draining (TdLN) and contralateral (cLN) lymph nodes were collected and assayed for VEGF-C levels by ELISA and for flow cytometry for (b) CD45.2 + /CD19 + B cells, and (c) CD45.2 + /CD11b + /F4/80 + macrophages. (d) Representative section of a WT sentinel lymph node showing B220 and VEGF-C co-staining (scale bar = 20 microns). (e) VEGF-C production by WT and A2aKO purified lymph node B cells. Means ± standard errors are shown. * p
Figure Legend Snippet: The number of VEGF-C secreting B cells is reduced in A2a-deficient sentinel lymph nodes. WT and A2a-deficient mice were injected with 2.5 × 10 5 B16-CD73+ melanoma cells in the foot pad. (a) Two weeks later, tumor-draining (TdLN) and contralateral (cLN) lymph nodes were collected and assayed for VEGF-C levels by ELISA and for flow cytometry for (b) CD45.2 + /CD19 + B cells, and (c) CD45.2 + /CD11b + /F4/80 + macrophages. (d) Representative section of a WT sentinel lymph node showing B220 and VEGF-C co-staining (scale bar = 20 microns). (e) VEGF-C production by WT and A2aKO purified lymph node B cells. Means ± standard errors are shown. * p

Techniques Used: Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Staining, Purification

19) Product Images from "VEGF-C promotes the development of lymphatics in bone and bone loss"

Article Title: VEGF-C promotes the development of lymphatics in bone and bone loss

Journal: eLife

doi: 10.7554/eLife.34323

Bone lymphatics in Osx-tTA;TetO-Vegfc mice disappear following the withdrawal of VEGF-C. ( A ) Schematic showing when mice received normal water and doxycycline water. One cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d). A second cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P38 (On 35d/Off 3d). A third cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P42 (On 35d/Off 7d). ( B ) Graph showing the relative VEGF-C mRNA levels in tibias from mice. ( C ) Schematic showing when mice received normal water and doxycycline water. Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d) or normal water from E18.5 to P35 and then doxycycline water for 3 (On 35d/Off 3d), 7 (On 35d/Off 7d), 28 (On 35d/Off 28d), or 56 days (On 35d/Off 56d). ( D–I ) Representative images of ribs stained with an anti-podoplanin antibody. The dashed lines separate the bone from the periosseous muscle. ( J ) Graph showing lymphatic vessel index values for ribs in Osx-tTA mice (0 ± 0.0; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (152.5 ± 29.56; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (62.25 ± 51.7; n = 4), 7 (24.08 ± 21.26; n = 4), 28 (0 ± 0.0; n = 5) or 56 (0 ± 0.0; n = 3) days. ( K ) Graph showing lymphatic vessel index values for periosseous muscle in Osx-tTA mice (3.61 ± 1.974; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (92.45 ± 34.63; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (99.29 ± 23.37; n = 4), 7 (74.84 ± 18.98; n = 4), 28 (92.67 ± 24.2; n = 5) or 56 (72.17 ± 14.05; n = 3) days. (**p
Figure Legend Snippet: Bone lymphatics in Osx-tTA;TetO-Vegfc mice disappear following the withdrawal of VEGF-C. ( A ) Schematic showing when mice received normal water and doxycycline water. One cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d). A second cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P38 (On 35d/Off 3d). A third cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P42 (On 35d/Off 7d). ( B ) Graph showing the relative VEGF-C mRNA levels in tibias from mice. ( C ) Schematic showing when mice received normal water and doxycycline water. Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d) or normal water from E18.5 to P35 and then doxycycline water for 3 (On 35d/Off 3d), 7 (On 35d/Off 7d), 28 (On 35d/Off 28d), or 56 days (On 35d/Off 56d). ( D–I ) Representative images of ribs stained with an anti-podoplanin antibody. The dashed lines separate the bone from the periosseous muscle. ( J ) Graph showing lymphatic vessel index values for ribs in Osx-tTA mice (0 ± 0.0; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (152.5 ± 29.56; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (62.25 ± 51.7; n = 4), 7 (24.08 ± 21.26; n = 4), 28 (0 ± 0.0; n = 5) or 56 (0 ± 0.0; n = 3) days. ( K ) Graph showing lymphatic vessel index values for periosseous muscle in Osx-tTA mice (3.61 ± 1.974; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (92.45 ± 34.63; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (99.29 ± 23.37; n = 4), 7 (74.84 ± 18.98; n = 4), 28 (92.67 ± 24.2; n = 5) or 56 (72.17 ± 14.05; n = 3) days. (**p

Techniques Used: Mouse Assay, Staining

Lymphatics in soft tissues in Osx-tTA;TetO-Vegfc mice appear normal. ( A ) Representative pictures of tissues collected from P35 Osx-tTA and Osx-tTA;TetO-Vegfc mice. Lymphatics in the lung, liver, kidney, and pancreas appeared normal in Osx-tTA;TetO-Vegfc mice. ( B ) Graph showing the circulating level of VEGF-C in P35 Osx-tTA (2.04 ± 1.42, n = 8) and Osx-tTA;TetO-Vegfc (2.387 ± 0.94, n = 7) mice.
Figure Legend Snippet: Lymphatics in soft tissues in Osx-tTA;TetO-Vegfc mice appear normal. ( A ) Representative pictures of tissues collected from P35 Osx-tTA and Osx-tTA;TetO-Vegfc mice. Lymphatics in the lung, liver, kidney, and pancreas appeared normal in Osx-tTA;TetO-Vegfc mice. ( B ) Graph showing the circulating level of VEGF-C in P35 Osx-tTA (2.04 ± 1.42, n = 8) and Osx-tTA;TetO-Vegfc (2.387 ± 0.94, n = 7) mice.

Techniques Used: Mouse Assay

Expression of VEGF-C during embryonic development causes embryonic lethality. ( A ) Schematic showing when mice received normal water. ( B–C ) Representative images of E12.5, E14.5 and E16.5 control embryos. ( E–G ) Representative images of E12.5, E14.5 and E16.5 Osx-tTA;TetO-Vegfc embryos. ( H,I ) Transverse sections of E14.5 embryos stained with hematoxylin and eosin. The jugular lymph sac (asterisk) in the control embryo appears normal. In contrast, the jugular lymph sac (asterisk) in the Osx-tTA;TetO-Vegfc embryo is enlarged.
Figure Legend Snippet: Expression of VEGF-C during embryonic development causes embryonic lethality. ( A ) Schematic showing when mice received normal water. ( B–C ) Representative images of E12.5, E14.5 and E16.5 control embryos. ( E–G ) Representative images of E12.5, E14.5 and E16.5 Osx-tTA;TetO-Vegfc embryos. ( H,I ) Transverse sections of E14.5 embryos stained with hematoxylin and eosin. The jugular lymph sac (asterisk) in the control embryo appears normal. In contrast, the jugular lymph sac (asterisk) in the Osx-tTA;TetO-Vegfc embryo is enlarged.

Techniques Used: Expressing, Mouse Assay, Staining

20) Product Images from "Effect of Niacin on Inflammation and Angiogenesis in a Murine Model of Ulcerative Colitis"

Article Title: Effect of Niacin on Inflammation and Angiogenesis in a Murine Model of Ulcerative Colitis

Journal: Scientific Reports

doi: 10.1038/s41598-017-07280-y

Effect of niacin on proangiogenic and antiangiogenic factors in colonic tissues of rats with iodoacetamide-induced colitis. (a) Vascular endothelial growth factor (VEGF), (b) angiostatin, (c) endostatin. Data are expressed as means ± SEM of 8 animals. # P ≤ 0.05 vs. normal control, * P ≤ 0.05 vs. colitis control.
Figure Legend Snippet: Effect of niacin on proangiogenic and antiangiogenic factors in colonic tissues of rats with iodoacetamide-induced colitis. (a) Vascular endothelial growth factor (VEGF), (b) angiostatin, (c) endostatin. Data are expressed as means ± SEM of 8 animals. # P ≤ 0.05 vs. normal control, * P ≤ 0.05 vs. colitis control.

Techniques Used:

21) Product Images from "Unexpected contribution of lymphatic vessels to promotion of distant metastatic tumor spread"

Article Title: Unexpected contribution of lymphatic vessels to promotion of distant metastatic tumor spread

Journal: Science Advances

doi: 10.1126/sciadv.aat4758

Characterization of CCSP-rtTA × tet-O–VEGF-C mice after doxycycline treatment for 2 weeks. ( A ) VEGF-C mRNA expression in the lung (normalized to RPLP0). The average expression level in WT × tet-O–VEGF-C mice was set to 1 ( n = 6). ( B ) VEGF-C protein levels in the lung detected by enzyme-linked immunosorbent assay (ELISA) ( n = 6). ( C ) Quantification of the VEGFR3 + LV area ( n = 4). ( D ) Representative images of VEGFR3 staining in the lung. Scale bars, 1 mm. ( E ) Representative high-magnification images of different regions in the lung stained for VEGFR3. Scale bars, 200 μm. PA, pulmonary artery; PV, pulmonary vein. ( F ) Representative images of MECA-32 staining in the lung. Scale bars, 50 μm. ( G ) Quantification of MECA-32 + blood vessel area in the lung ( n = 5).
Figure Legend Snippet: Characterization of CCSP-rtTA × tet-O–VEGF-C mice after doxycycline treatment for 2 weeks. ( A ) VEGF-C mRNA expression in the lung (normalized to RPLP0). The average expression level in WT × tet-O–VEGF-C mice was set to 1 ( n = 6). ( B ) VEGF-C protein levels in the lung detected by enzyme-linked immunosorbent assay (ELISA) ( n = 6). ( C ) Quantification of the VEGFR3 + LV area ( n = 4). ( D ) Representative images of VEGFR3 staining in the lung. Scale bars, 1 mm. ( E ) Representative high-magnification images of different regions in the lung stained for VEGFR3. Scale bars, 200 μm. PA, pulmonary artery; PV, pulmonary vein. ( F ) Representative images of MECA-32 staining in the lung. Scale bars, 50 μm. ( G ) Quantification of MECA-32 + blood vessel area in the lung ( n = 5).

Techniques Used: Mouse Assay, Expressing, Enzyme-linked Immunosorbent Assay, Staining

Increased metastasis in CCSP-rtTA × tet-O–VEGF-C mice after tail vein injection of B16F10 melanoma cells. ( A ) Schematic of the B16F10 tumor study. Dox, doxycycline; i.v., intravenous. ( B ) Representative images of lung sections with metastatic nodules 14 days after tumor cell injection (indicated by staining for RFP from B16F10-luc2-tdTomato cells). Scale bars, 1 mm. ( C ) Quantification of tdTomato + area in the lung 14 days after tumor cell injection ( n = 10). ( D ) Quantification of tdTomato + cells per lung 24 hours after intravenous injection of 1 × 10 6 B16F10-luc2-tdTomato cells. ( E ) Representative images of lymphatic invasion (white arrows) with orthogonal projection. Scale bars, 100 μm. Fisher’s exact test was performed. ( F ) Quantification of the lung-draining lymph node weight ( n = 10). ( G ) Number of mice with different numbers of distant organs with metastasis (zero, one, two, or three organs affected) and example images of metastases (yellow arrows) in the liver and intestine. Data were pooled from two rounds of studies.
Figure Legend Snippet: Increased metastasis in CCSP-rtTA × tet-O–VEGF-C mice after tail vein injection of B16F10 melanoma cells. ( A ) Schematic of the B16F10 tumor study. Dox, doxycycline; i.v., intravenous. ( B ) Representative images of lung sections with metastatic nodules 14 days after tumor cell injection (indicated by staining for RFP from B16F10-luc2-tdTomato cells). Scale bars, 1 mm. ( C ) Quantification of tdTomato + area in the lung 14 days after tumor cell injection ( n = 10). ( D ) Quantification of tdTomato + cells per lung 24 hours after intravenous injection of 1 × 10 6 B16F10-luc2-tdTomato cells. ( E ) Representative images of lymphatic invasion (white arrows) with orthogonal projection. Scale bars, 100 μm. Fisher’s exact test was performed. ( F ) Quantification of the lung-draining lymph node weight ( n = 10). ( G ) Number of mice with different numbers of distant organs with metastasis (zero, one, two, or three organs affected) and example images of metastases (yellow arrows) in the liver and intestine. Data were pooled from two rounds of studies.

Techniques Used: Mouse Assay, Injection, Staining

Increased metastasis in CCSP-rtTA × tet-O–VEGF-C mice in the 4T1 spontaneous breast cancer metastasis model. ( A ) Schematic of the 4T1 tumor study. PR, postremoval; s.c., subcutaneous. ( B ) Representative in vivo bioluminescence images of metastases on day 21 after primary tumor removal. ( C ) Quantification of bioluminescence signal over the whole body on days PR7, PR14, and PR21 by IVIS imaging. For data from days PR7 and PR14, WT × VEGFC, n = 8, and CCSP × VEGFC, n = 11. Four mice reached the euthanization criteria before the end time point; thus, for data from day PR 21, WT × VEGFC, n = 7, and CCSP × VEGFC, n = 8. ns, not significant. ( D ) Number of mice with different numbers of distant organs with metastasis (zero, one, two, or three organs affected). Data were pooled from three rounds of studies. Only mice with detectable metastases (by ex vivo IVIS imaging) in the lungs were quantified.
Figure Legend Snippet: Increased metastasis in CCSP-rtTA × tet-O–VEGF-C mice in the 4T1 spontaneous breast cancer metastasis model. ( A ) Schematic of the 4T1 tumor study. PR, postremoval; s.c., subcutaneous. ( B ) Representative in vivo bioluminescence images of metastases on day 21 after primary tumor removal. ( C ) Quantification of bioluminescence signal over the whole body on days PR7, PR14, and PR21 by IVIS imaging. For data from days PR7 and PR14, WT × VEGFC, n = 8, and CCSP × VEGFC, n = 11. Four mice reached the euthanization criteria before the end time point; thus, for data from day PR 21, WT × VEGFC, n = 7, and CCSP × VEGFC, n = 8. ns, not significant. ( D ) Number of mice with different numbers of distant organs with metastasis (zero, one, two, or three organs affected). Data were pooled from three rounds of studies. Only mice with detectable metastases (by ex vivo IVIS imaging) in the lungs were quantified.

Techniques Used: Mouse Assay, In Vivo, Imaging, Ex Vivo

22) Product Images from "Effects of propofol/remifentanil-based total intravenous anesthesia versus sevoflurane-based inhalational anesthesia on the release of VEGF-C and TGF-β and prognosis after breast cancer surgery: a prospective, randomized and controlled study"

Article Title: Effects of propofol/remifentanil-based total intravenous anesthesia versus sevoflurane-based inhalational anesthesia on the release of VEGF-C and TGF-β and prognosis after breast cancer surgery: a prospective, randomized and controlled study

Journal: BMC Anesthesiology

doi: 10.1186/s12871-018-0588-3

Median preoperative and postoperative VEGF-C concentrations in both groups. * P = 0.009,higher postoperative values versus preoperative values in the SEV group. SEV-pre preoperative values in the SEV group, SEV-post postoperative values in the SEV group, TIVA-pre preoperative values in the SEV group, TIVA-post postoperative values in the TIVA group. Horizontal line denotes median values, box borders refer to interquartile range, whiskers indicate range of values
Figure Legend Snippet: Median preoperative and postoperative VEGF-C concentrations in both groups. * P = 0.009,higher postoperative values versus preoperative values in the SEV group. SEV-pre preoperative values in the SEV group, SEV-post postoperative values in the SEV group, TIVA-pre preoperative values in the SEV group, TIVA-post postoperative values in the TIVA group. Horizontal line denotes median values, box borders refer to interquartile range, whiskers indicate range of values

Techniques Used:

23) Product Images from "Tissue factor induces VEGF expression via activation of the Wnt/β-catenin signaling pathway in ARPE-19 cells"

Article Title: Tissue factor induces VEGF expression via activation of the Wnt/β-catenin signaling pathway in ARPE-19 cells

Journal: Molecular Vision

doi:

Chemical hypoxia induces TF, HIF-1α, and VEGF expression in ARPE-19 cells. A : Western blots show tissue factor (TF) and hypoxia-inducible factor 1-alpha (HIF-1α). B : Histogram shows the densitometric analysis of the average levels for TF and HIF-1α to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in each group. C : The vascular endothelial growth factor (VEGF) concentrations are examined with enzyme-linked immunosorbent assay (ELISA) at different hypoxia time points. Hypoxia significantly increased the TF, HIF-1α, and VEGF levels in the ARPE-19 cells. *p
Figure Legend Snippet: Chemical hypoxia induces TF, HIF-1α, and VEGF expression in ARPE-19 cells. A : Western blots show tissue factor (TF) and hypoxia-inducible factor 1-alpha (HIF-1α). B : Histogram shows the densitometric analysis of the average levels for TF and HIF-1α to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in each group. C : The vascular endothelial growth factor (VEGF) concentrations are examined with enzyme-linked immunosorbent assay (ELISA) at different hypoxia time points. Hypoxia significantly increased the TF, HIF-1α, and VEGF levels in the ARPE-19 cells. *p

Techniques Used: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

24) Product Images from "Biological effects of dosing aerobic exercise and neuromuscular electrical stimulation in rats"

Article Title: Biological effects of dosing aerobic exercise and neuromuscular electrical stimulation in rats

Journal: Scientific Reports

doi: 10.1038/s41598-017-11260-7

Contour maps – systemic factors. Based on the factorial design that arrayed treatment interventions to a control (0 dose), low, medium and high dose, contour plots of the dependent variables can be drawn to illustrate how these variables interact across the experimental space. It is evident here that systemic BDNF levels were most dramatically increased using NMES, but due to the lack of increase in MCT performance there was little interaction between treatment dose and % change in speed. BDNF did also not reveal a dramatic dose-dependent interaction for AE. A similar lack of interaction was evident for VEGF-A, with some evidence of an increase in levels for a low AE condition that results in a 25–50% change in speed. A clearer interaction between dosing and MCT was evident for AE on IGF-1. A low to medium dose of therapy here produced higher levels of factor release, but this was associated with a poorer performance. Lower levels of IGF-1 were actually induced by the high AE condition, producing the most significant performance change (upper right corner). Minimal interaction was seen for NMES, considering that there was less of an effect on MCT, demonstrating that IGF-1 levels in this paradigm were almost homogenous throughout experimental space. In the AE condition, the contour plot for Klotho indicated that the medium dose produced the highest level of Klotho release, which was associated with the highest level of behavioral change. In contrast the plot for NMES did not reveal a clear interaction between these variables.
Figure Legend Snippet: Contour maps – systemic factors. Based on the factorial design that arrayed treatment interventions to a control (0 dose), low, medium and high dose, contour plots of the dependent variables can be drawn to illustrate how these variables interact across the experimental space. It is evident here that systemic BDNF levels were most dramatically increased using NMES, but due to the lack of increase in MCT performance there was little interaction between treatment dose and % change in speed. BDNF did also not reveal a dramatic dose-dependent interaction for AE. A similar lack of interaction was evident for VEGF-A, with some evidence of an increase in levels for a low AE condition that results in a 25–50% change in speed. A clearer interaction between dosing and MCT was evident for AE on IGF-1. A low to medium dose of therapy here produced higher levels of factor release, but this was associated with a poorer performance. Lower levels of IGF-1 were actually induced by the high AE condition, producing the most significant performance change (upper right corner). Minimal interaction was seen for NMES, considering that there was less of an effect on MCT, demonstrating that IGF-1 levels in this paradigm were almost homogenous throughout experimental space. In the AE condition, the contour plot for Klotho indicated that the medium dose produced the highest level of Klotho release, which was associated with the highest level of behavioral change. In contrast the plot for NMES did not reveal a clear interaction between these variables.

Techniques Used: Produced

Comparison of systemic factors using enzyme-linked immunosorbent assays (ELISA). Brain-derived nerve growth factor (BDNF) was upregulated after both AE and NMES, with the most dramatic change evident after a medium dose of NMES resulting in an almost 8-fold increase. Little change was evident in VEGF-A, with only the low AE condition showing a minor non-significant 6% increase. The low AE condition also increased IGF-1 by 33% and showed an AE dose-dependent decrease, with the medium condition being equivalent to control levels. NMES did not affect IGF-1 levels. In contrast, Klotho revealed a significant AE dose-dependent modulation of levels with a medium dose of AE doubling the amount of Klotho in the blood compared to controls. NMES also produced an increase of Klotho for the low and high condition, but not for the medium dose. (*p
Figure Legend Snippet: Comparison of systemic factors using enzyme-linked immunosorbent assays (ELISA). Brain-derived nerve growth factor (BDNF) was upregulated after both AE and NMES, with the most dramatic change evident after a medium dose of NMES resulting in an almost 8-fold increase. Little change was evident in VEGF-A, with only the low AE condition showing a minor non-significant 6% increase. The low AE condition also increased IGF-1 by 33% and showed an AE dose-dependent decrease, with the medium condition being equivalent to control levels. NMES did not affect IGF-1 levels. In contrast, Klotho revealed a significant AE dose-dependent modulation of levels with a medium dose of AE doubling the amount of Klotho in the blood compared to controls. NMES also produced an increase of Klotho for the low and high condition, but not for the medium dose. (*p

Techniques Used: Enzyme-linked Immunosorbent Assay, Derivative Assay, Produced

Hippocampal gene expression. BDNF mRNA was upregulated after AE, but did not exhibit a dose-dependency. A significant 10% increase in hippocampal BDNF was evident after low NMES and showed a dose dependent decrease to the level of controls. VEGF-A was upregulated in all NMES conditions, but did not change after a medium and high dose of AE. A low dose actually decreased VEGF-A expression. IGF-1 was upregulated in all conditions by approx. 17%. Only the low AE condition produced a lower 5% upregulation. Klotho revealed an expression profile similar to IGF-1 (27% increase), but with a more marked increase in expression in the low AE condition (12%). (*p
Figure Legend Snippet: Hippocampal gene expression. BDNF mRNA was upregulated after AE, but did not exhibit a dose-dependency. A significant 10% increase in hippocampal BDNF was evident after low NMES and showed a dose dependent decrease to the level of controls. VEGF-A was upregulated in all NMES conditions, but did not change after a medium and high dose of AE. A low dose actually decreased VEGF-A expression. IGF-1 was upregulated in all conditions by approx. 17%. Only the low AE condition produced a lower 5% upregulation. Klotho revealed an expression profile similar to IGF-1 (27% increase), but with a more marked increase in expression in the low AE condition (12%). (*p

Techniques Used: Expressing, Produced

Contour maps – hippocampal gene expression. By accounting for the different levels of dosing for AE and NMES, contour maps were generated to expose interactions between variables in the experimental space. It was evident here that BDNF expression in the hippocampus was highest for the high AE and low NMES conditions. As there was no significant improvement in MCT performance from NMES, this produced a rather flat banding of expression levels. The highest BDNF levels were evident between no change to a 25% performance decrease. For AE, the highest levels were achieved by a high dose that resulted in the highest level of performance increase. VEGF-A revealed the highest increase in the NMES group with a low to a medium dosing leading to peak gene expression. This was associated with improvements in MCT. AE yielded a dose-dependent increase in IGF-1 signaling that was linked to improved performance with a diagonal pattern across the experimental space. A similar effect was evident for Klotho with lower levels being associated with poorer performance and higher levels indicating better performance. A medium dosing of AE is sufficient to achieve peak performance and high levels of Klotho. It was also highly upregulated in the NMES condition with only very high decreases in performance showing a low level of Klotho.
Figure Legend Snippet: Contour maps – hippocampal gene expression. By accounting for the different levels of dosing for AE and NMES, contour maps were generated to expose interactions between variables in the experimental space. It was evident here that BDNF expression in the hippocampus was highest for the high AE and low NMES conditions. As there was no significant improvement in MCT performance from NMES, this produced a rather flat banding of expression levels. The highest BDNF levels were evident between no change to a 25% performance decrease. For AE, the highest levels were achieved by a high dose that resulted in the highest level of performance increase. VEGF-A revealed the highest increase in the NMES group with a low to a medium dosing leading to peak gene expression. This was associated with improvements in MCT. AE yielded a dose-dependent increase in IGF-1 signaling that was linked to improved performance with a diagonal pattern across the experimental space. A similar effect was evident for Klotho with lower levels being associated with poorer performance and higher levels indicating better performance. A medium dosing of AE is sufficient to achieve peak performance and high levels of Klotho. It was also highly upregulated in the NMES condition with only very high decreases in performance showing a low level of Klotho.

Techniques Used: Expressing, Generated, Produced

25) Product Images from "VEGF-C promotes the development of lymphatics in bone and bone loss"

Article Title: VEGF-C promotes the development of lymphatics in bone and bone loss

Journal: eLife

doi: 10.7554/eLife.34323

Bone lymphatics in Osx-tTA;TetO-Vegfc mice disappear following the withdrawal of VEGF-C. ( A ) Schematic showing when mice received normal water and doxycycline water. One cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d). A second cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P38 (On 35d/Off 3d). A third cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P42 (On 35d/Off 7d). ( B ) Graph showing the relative VEGF-C mRNA levels in tibias from mice. ( C ) Schematic showing when mice received normal water and doxycycline water. Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d) or normal water from E18.5 to P35 and then doxycycline water for 3 (On 35d/Off 3d), 7 (On 35d/Off 7d), 28 (On 35d/Off 28d), or 56 days (On 35d/Off 56d). ( D–I ) Representative images of ribs stained with an anti-podoplanin antibody. The dashed lines separate the bone from the periosseous muscle. ( J ) Graph showing lymphatic vessel index values for ribs in Osx-tTA mice (0 ± 0.0; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (152.5 ± 29.56; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (62.25 ± 51.7; n = 4), 7 (24.08 ± 21.26; n = 4), 28 (0 ± 0.0; n = 5) or 56 (0 ± 0.0; n = 3) days. ( K ) Graph showing lymphatic vessel index values for periosseous muscle in Osx-tTA mice (3.61 ± 1.974; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (92.45 ± 34.63; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (99.29 ± 23.37; n = 4), 7 (74.84 ± 18.98; n = 4), 28 (92.67 ± 24.2; n = 5) or 56 (72.17 ± 14.05; n = 3) days. (**p
Figure Legend Snippet: Bone lymphatics in Osx-tTA;TetO-Vegfc mice disappear following the withdrawal of VEGF-C. ( A ) Schematic showing when mice received normal water and doxycycline water. One cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d). A second cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P38 (On 35d/Off 3d). A third cohort of Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 and then doxycycline water from P35 to P42 (On 35d/Off 7d). ( B ) Graph showing the relative VEGF-C mRNA levels in tibias from mice. ( C ) Schematic showing when mice received normal water and doxycycline water. Osx-tTA;TetO-Vegfc mice received normal water from E18.5 to P35 (On 35d) or normal water from E18.5 to P35 and then doxycycline water for 3 (On 35d/Off 3d), 7 (On 35d/Off 7d), 28 (On 35d/Off 28d), or 56 days (On 35d/Off 56d). ( D–I ) Representative images of ribs stained with an anti-podoplanin antibody. The dashed lines separate the bone from the periosseous muscle. ( J ) Graph showing lymphatic vessel index values for ribs in Osx-tTA mice (0 ± 0.0; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (152.5 ± 29.56; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (62.25 ± 51.7; n = 4), 7 (24.08 ± 21.26; n = 4), 28 (0 ± 0.0; n = 5) or 56 (0 ± 0.0; n = 3) days. ( K ) Graph showing lymphatic vessel index values for periosseous muscle in Osx-tTA mice (3.61 ± 1.974; n = 5), Osx-tTA;TetO-Vegfc mice that received normal water for 35 days (92.45 ± 34.63; n = 5), and Osx-tTA;TetO-Vegfc mice that received normal water for 35 days and then doxycycline water for 3 (99.29 ± 23.37; n = 4), 7 (74.84 ± 18.98; n = 4), 28 (92.67 ± 24.2; n = 5) or 56 (72.17 ± 14.05; n = 3) days. (**p

Techniques Used: Mouse Assay, Staining

Osx-tTA;TetO-Vegfc mice develop lymphatics in their bones. ( A ) Schematic of the Tet-Off system used to express VEGF-C in bone. Doxycycline inhibits the expression of VEGF-C. ( B ) Schematic showing when mice received normal water and doxycycline water. Osx-tTA and Osx-tTA;TetO-Vegfc mice received doxycycline water from E0.5 to E18.5 and then normal water from E18.5 to P35. ( C–H ) Representative images of cortical bone in Osx-tTA and Osx-tTA;TetO-Vegfc femurs. Sections were stained with an anti-podoplanin antibody. Arrowheads point to podoplanin-positive osteocytes. Arrows point to podoplanin-positive lymphatics. ( I ) Graph showing lymphatic vessel index values for cortical bone in P21 (0 ± 0.0; n = 3), P28 (0 ± 0.0; n = 5), and P35 (0 ± 0.0; n = 6) Osx-tTA mice and in P21 (0 ± 0.0; n = 4), P28 (67 ± 22.06; n = 4), and P35 (130.3 ± 44.35; n = 4) Osx-tTA;TetO-Vegfc mice. ( J–O ) Representative images of trabecular bone in Osx-tTA and Osx-tTA;TetO-Vegfc femurs. Sections were stained with an anti-podoplanin antibody. Arrowheads point to podoplanin-positive osteocytes. Arrow points to podoplanin-positive lymphatics. ( P ) Graph showing lymphatic vessel index values for trabecular bone in P21 (0 ± 0.0; n = 3), P28 (0 ± 0.0; n = 5), and P35 (0 ± 0.0; n = 6) Osx-tTA mice and P21 (0 ± 0.0; n = 4), P28 (0.167 ± 0.4082; n = 6), and P35 (189.5 ± 47.7; n = 4) Osx-tTA;TetO-Vegfc mice. (***p
Figure Legend Snippet: Osx-tTA;TetO-Vegfc mice develop lymphatics in their bones. ( A ) Schematic of the Tet-Off system used to express VEGF-C in bone. Doxycycline inhibits the expression of VEGF-C. ( B ) Schematic showing when mice received normal water and doxycycline water. Osx-tTA and Osx-tTA;TetO-Vegfc mice received doxycycline water from E0.5 to E18.5 and then normal water from E18.5 to P35. ( C–H ) Representative images of cortical bone in Osx-tTA and Osx-tTA;TetO-Vegfc femurs. Sections were stained with an anti-podoplanin antibody. Arrowheads point to podoplanin-positive osteocytes. Arrows point to podoplanin-positive lymphatics. ( I ) Graph showing lymphatic vessel index values for cortical bone in P21 (0 ± 0.0; n = 3), P28 (0 ± 0.0; n = 5), and P35 (0 ± 0.0; n = 6) Osx-tTA mice and in P21 (0 ± 0.0; n = 4), P28 (67 ± 22.06; n = 4), and P35 (130.3 ± 44.35; n = 4) Osx-tTA;TetO-Vegfc mice. ( J–O ) Representative images of trabecular bone in Osx-tTA and Osx-tTA;TetO-Vegfc femurs. Sections were stained with an anti-podoplanin antibody. Arrowheads point to podoplanin-positive osteocytes. Arrow points to podoplanin-positive lymphatics. ( P ) Graph showing lymphatic vessel index values for trabecular bone in P21 (0 ± 0.0; n = 3), P28 (0 ± 0.0; n = 5), and P35 (0 ± 0.0; n = 6) Osx-tTA mice and P21 (0 ± 0.0; n = 4), P28 (0.167 ± 0.4082; n = 6), and P35 (189.5 ± 47.7; n = 4) Osx-tTA;TetO-Vegfc mice. (***p

Techniques Used: Mouse Assay, Expressing, Staining

26) Product Images from "Therapeutic Lymphangiogenesis With Implantation of Adipose-Derived Regenerative Cells"

Article Title: Therapeutic Lymphangiogenesis With Implantation of Adipose-Derived Regenerative Cells

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

doi: 10.1161/JAHA.112.000877

The ability of ADRCs for lymphangiogenesis in vitro (functional assay and differentiation assay). A, Enzyme‐linked immunosorbent assay (ELISA) revealed that the concentration of VEGF‐C was upregulated in ADRC‐CM compared to control. B, rhVEGF‐C induced LEC migration in a dose‐dependent manner, and ADRC‐CM also induced LEC migration. HPF indicates high‐powered field. * P
Figure Legend Snippet: The ability of ADRCs for lymphangiogenesis in vitro (functional assay and differentiation assay). A, Enzyme‐linked immunosorbent assay (ELISA) revealed that the concentration of VEGF‐C was upregulated in ADRC‐CM compared to control. B, rhVEGF‐C induced LEC migration in a dose‐dependent manner, and ADRC‐CM also induced LEC migration. HPF indicates high‐powered field. * P

Techniques Used: In Vitro, Functional Assay, Differentiation Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Migration

27) Product Images from "Combination leflunomide and methotrexate impedes the recovery of liver fibrosis, partly through inhibition of myeloid cell admittance"

Article Title: Combination leflunomide and methotrexate impedes the recovery of liver fibrosis, partly through inhibition of myeloid cell admittance

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2019.9821

Serum collagen is sustained at high levels in mice treated with LEF+MTX. The serum was collected at weeks 2 and 4 during the recovery phase. The levels of (A) collagen IV, (B) hyaluronic acid, (C) laminin and (D) procollagen III in serum were measured by ELISA. Data are presented as the mean ± standard deviation (n=6). *P
Figure Legend Snippet: Serum collagen is sustained at high levels in mice treated with LEF+MTX. The serum was collected at weeks 2 and 4 during the recovery phase. The levels of (A) collagen IV, (B) hyaluronic acid, (C) laminin and (D) procollagen III in serum were measured by ELISA. Data are presented as the mean ± standard deviation (n=6). *P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

28) Product Images from "Ethyl-p-methoxycinnamate isolated from kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-?, and angiogenesis by blocking endothelial functions"

Article Title: Ethyl-p-methoxycinnamate isolated from kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-?, and angiogenesis by blocking endothelial functions

Journal: Clinics

doi: 10.6061/clinics/2014(02)10

Enzyme-linked immunosorbant assay to measure TNF-α and IL-1
Figure Legend Snippet: Enzyme-linked immunosorbant assay to measure TNF-α and IL-1

Techniques Used:

A ) The mean weight of cotton pellets dissected from rats treated with ethyl- p -methoxycinnamate (EPMC) at doses of 200, 400, and 800 mg/kg. B ) Percent increase in the time taken by rats to flick their tail away from heat, a measure of the analgesic effect of EPMC, at doses of 200, 400, and 800 mg/kg. C ) Blood concentration of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) in rats treated with EPMC at doses of 200, 400, and 800 mg/kg on the 8 th day of the cotton pellet granuloma assay. D ) Percent inhibition of IL-1, TNF-α, and nitric oxide (NO) synthesis by EPMC in U937 cells at concentrations of 25, 50, 100, and 200 µg/ml. The cell culture supernatants were collected after 48 hours and subjected to an enzyme-linked immunosorbant assay (ELISA) for the assessment of IL-1, TNF-α, and NO. All values are represented as the mean ± SEM ( n = 6), and * , ** , and *** indicate significant differences of p
Figure Legend Snippet: A ) The mean weight of cotton pellets dissected from rats treated with ethyl- p -methoxycinnamate (EPMC) at doses of 200, 400, and 800 mg/kg. B ) Percent increase in the time taken by rats to flick their tail away from heat, a measure of the analgesic effect of EPMC, at doses of 200, 400, and 800 mg/kg. C ) Blood concentration of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) in rats treated with EPMC at doses of 200, 400, and 800 mg/kg on the 8 th day of the cotton pellet granuloma assay. D ) Percent inhibition of IL-1, TNF-α, and nitric oxide (NO) synthesis by EPMC in U937 cells at concentrations of 25, 50, 100, and 200 µg/ml. The cell culture supernatants were collected after 48 hours and subjected to an enzyme-linked immunosorbant assay (ELISA) for the assessment of IL-1, TNF-α, and NO. All values are represented as the mean ± SEM ( n = 6), and * , ** , and *** indicate significant differences of p

Techniques Used: Concentration Assay, Inhibition, Cell Culture, Enzyme-linked Immunosorbent Assay

29) Product Images from "Unexpected contribution of lymphatic vessels to promotion of distant metastatic tumor spread"

Article Title: Unexpected contribution of lymphatic vessels to promotion of distant metastatic tumor spread

Journal: Science Advances

doi: 10.1126/sciadv.aat4758

Characterization of CCSP-rtTA × tet-O–VEGF-C mice after doxycycline treatment for 2 weeks. ( A ) VEGF-C mRNA expression in the lung (normalized to RPLP0). The average expression level in WT × tet-O–VEGF-C mice was set to 1 ( n = 6). ( B ) VEGF-C protein levels in the lung detected by enzyme-linked immunosorbent assay (ELISA) ( n = 6). ( C ) Quantification of the VEGFR3 + LV area ( n = 4). ( D ) Representative images of VEGFR3 staining in the lung. Scale bars, 1 mm. ( E ) Representative high-magnification images of different regions in the lung stained for VEGFR3. Scale bars, 200 μm. PA, pulmonary artery; PV, pulmonary vein. ( F ) Representative images of MECA-32 staining in the lung. Scale bars, 50 μm. ( G ) Quantification of MECA-32 + blood vessel area in the lung ( n = 5).
Figure Legend Snippet: Characterization of CCSP-rtTA × tet-O–VEGF-C mice after doxycycline treatment for 2 weeks. ( A ) VEGF-C mRNA expression in the lung (normalized to RPLP0). The average expression level in WT × tet-O–VEGF-C mice was set to 1 ( n = 6). ( B ) VEGF-C protein levels in the lung detected by enzyme-linked immunosorbent assay (ELISA) ( n = 6). ( C ) Quantification of the VEGFR3 + LV area ( n = 4). ( D ) Representative images of VEGFR3 staining in the lung. Scale bars, 1 mm. ( E ) Representative high-magnification images of different regions in the lung stained for VEGFR3. Scale bars, 200 μm. PA, pulmonary artery; PV, pulmonary vein. ( F ) Representative images of MECA-32 staining in the lung. Scale bars, 50 μm. ( G ) Quantification of MECA-32 + blood vessel area in the lung ( n = 5).

Techniques Used: Mouse Assay, Expressing, Enzyme-linked Immunosorbent Assay, Staining

30) Product Images from "miR-129-5p targets Wnt5a to block PKC/ERK/NF-κB and JNK pathways in glioblastoma"

Article Title: miR-129-5p targets Wnt5a to block PKC/ERK/NF-κB and JNK pathways in glioblastoma

Journal: Cell Death & Disease

doi: 10.1038/s41419-018-0343-1

miR-129-5p overexpression inhibits cell proliferation and angiogenesis in GBM by targeting Wnt5a . a N3/miR-129-5p, K3/miR-129-5p and U251/miR-129-5p cells were transfected with the Wnt5a-plasmid vector, followed by western blot analysis of Wnt5a transcripts (three replicates per group, three independent experiments per group). GAPDH served as the loading control. b Colony formation ability of the miR-NC- or miR-129-5p-transfected N3 or U251 cells without transfection or transfected with the pcDNA3.1-Wnt5a plasmid (Wnt5a) (six replicates per group, three independent experiments per group). Scale bar, 2.5 mm. c Overexpression of miR-129-5p arrested cell proliferation; however, this was rescued upon coexpression of exogenous Wnt5a in N3 and U251 cells (six replicates per group, three independent experiments per group). d EdU analysis of miR-NC, miR-129-5p or miR-129-5p plus Wnt5a-transfected N3 and U251 cells (six replicates per group, three independent experiments per group). Scale bar, 100 μm. e Cell-cycle assay of N3 and U251 GBM cells 3 days after transfection with miR-NC, miR-129-5p or miR-129-5p plus Wnt5a (six replicates per group, three independent experiments per group). f Western blot analysis indicated the regulation of the cell-cycle-regulatory proteins cyclin E1 and cyclin D1 in miR-NC, miR-129-5p or miR-129-5p mimic plus Wnt5a-transfected N3 or U251 cells. GAPDH was used as the loading control (three replicates per group, three independent experiments per group). g Representative images and quantification of HBMVECs cultured on Matrigel-coated 96-well plates using conditioned medium from the indicated cells (six replicates per group, three independent experiments per group). Scale bar, 200 μm. Data are expressed as the mean ± s.e.m. h – i Quantitative determination of Wnt5a and VEGF levels in supernatants from N3 or U251 cells with indicated treatment by ELISA. (six replicates per group, three independent experiments per group). Data are expressed as the mean ± s.e.m
Figure Legend Snippet: miR-129-5p overexpression inhibits cell proliferation and angiogenesis in GBM by targeting Wnt5a . a N3/miR-129-5p, K3/miR-129-5p and U251/miR-129-5p cells were transfected with the Wnt5a-plasmid vector, followed by western blot analysis of Wnt5a transcripts (three replicates per group, three independent experiments per group). GAPDH served as the loading control. b Colony formation ability of the miR-NC- or miR-129-5p-transfected N3 or U251 cells without transfection or transfected with the pcDNA3.1-Wnt5a plasmid (Wnt5a) (six replicates per group, three independent experiments per group). Scale bar, 2.5 mm. c Overexpression of miR-129-5p arrested cell proliferation; however, this was rescued upon coexpression of exogenous Wnt5a in N3 and U251 cells (six replicates per group, three independent experiments per group). d EdU analysis of miR-NC, miR-129-5p or miR-129-5p plus Wnt5a-transfected N3 and U251 cells (six replicates per group, three independent experiments per group). Scale bar, 100 μm. e Cell-cycle assay of N3 and U251 GBM cells 3 days after transfection with miR-NC, miR-129-5p or miR-129-5p plus Wnt5a (six replicates per group, three independent experiments per group). f Western blot analysis indicated the regulation of the cell-cycle-regulatory proteins cyclin E1 and cyclin D1 in miR-NC, miR-129-5p or miR-129-5p mimic plus Wnt5a-transfected N3 or U251 cells. GAPDH was used as the loading control (three replicates per group, three independent experiments per group). g Representative images and quantification of HBMVECs cultured on Matrigel-coated 96-well plates using conditioned medium from the indicated cells (six replicates per group, three independent experiments per group). Scale bar, 200 μm. Data are expressed as the mean ± s.e.m. h – i Quantitative determination of Wnt5a and VEGF levels in supernatants from N3 or U251 cells with indicated treatment by ELISA. (six replicates per group, three independent experiments per group). Data are expressed as the mean ± s.e.m

Techniques Used: Over Expression, Transfection, Plasmid Preparation, Western Blot, Cell Cycle Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

31) Product Images from "Ethyl-p-methoxycinnamate isolated from kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-?, and angiogenesis by blocking endothelial functions"

Article Title: Ethyl-p-methoxycinnamate isolated from kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-?, and angiogenesis by blocking endothelial functions

Journal: Clinics

doi: 10.6061/clinics/2014(02)10

Enzyme-linked immunosorbant assay to measure TNF-α and IL-1
Figure Legend Snippet: Enzyme-linked immunosorbant assay to measure TNF-α and IL-1

Techniques Used:

A ) The mean weight of cotton pellets dissected from rats treated with ethyl- p -methoxycinnamate (EPMC) at doses of 200, 400, and 800 mg/kg. B ) Percent increase in the time taken by rats to flick their tail away from heat, a measure of the analgesic effect of EPMC, at doses of 200, 400, and 800 mg/kg. C ) Blood concentration of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) in rats treated with EPMC at doses of 200, 400, and 800 mg/kg on the 8 th day of the cotton pellet granuloma assay. D ) Percent inhibition of IL-1, TNF-α, and nitric oxide (NO) synthesis by EPMC in U937 cells at concentrations of 25, 50, 100, and 200 µg/ml. The cell culture supernatants were collected after 48 hours and subjected to an enzyme-linked immunosorbant assay (ELISA) for the assessment of IL-1, TNF-α, and NO. All values are represented as the mean ± SEM ( n = 6), and * , ** , and *** indicate significant differences of p
Figure Legend Snippet: A ) The mean weight of cotton pellets dissected from rats treated with ethyl- p -methoxycinnamate (EPMC) at doses of 200, 400, and 800 mg/kg. B ) Percent increase in the time taken by rats to flick their tail away from heat, a measure of the analgesic effect of EPMC, at doses of 200, 400, and 800 mg/kg. C ) Blood concentration of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) in rats treated with EPMC at doses of 200, 400, and 800 mg/kg on the 8 th day of the cotton pellet granuloma assay. D ) Percent inhibition of IL-1, TNF-α, and nitric oxide (NO) synthesis by EPMC in U937 cells at concentrations of 25, 50, 100, and 200 µg/ml. The cell culture supernatants were collected after 48 hours and subjected to an enzyme-linked immunosorbant assay (ELISA) for the assessment of IL-1, TNF-α, and NO. All values are represented as the mean ± SEM ( n = 6), and * , ** , and *** indicate significant differences of p

Techniques Used: Concentration Assay, Inhibition, Cell Culture, Enzyme-linked Immunosorbent Assay

32) Product Images from "Combination leflunomide and methotrexate impedes the recovery of liver fibrosis, partly through inhibition of myeloid cell admittance"

Article Title: Combination leflunomide and methotrexate impedes the recovery of liver fibrosis, partly through inhibition of myeloid cell admittance

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2019.9821

Serum collagen is sustained at high levels in mice treated with LEF+MTX. The serum was collected at weeks 2 and 4 during the recovery phase. The levels of (A) collagen IV, (B) hyaluronic acid, (C) laminin and (D) procollagen III in serum were measured by ELISA. Data are presented as the mean ± standard deviation (n=6). *P
Figure Legend Snippet: Serum collagen is sustained at high levels in mice treated with LEF+MTX. The serum was collected at weeks 2 and 4 during the recovery phase. The levels of (A) collagen IV, (B) hyaluronic acid, (C) laminin and (D) procollagen III in serum were measured by ELISA. Data are presented as the mean ± standard deviation (n=6). *P

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

33) Product Images from "Bone mesenchymal stem cells ameliorate ischemia/reperfusion-induced damage in renal epithelial cells via microRNA-223"

Article Title: Bone mesenchymal stem cells ameliorate ischemia/reperfusion-induced damage in renal epithelial cells via microRNA-223

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-017-0599-x

Knockdown of miR-223 blocked the protective effect of mesenchymal stem cells ( MSCs ) on renal tubular epithelial cells ( RTECs ). After the kidney ischemia/reperfusion ( IR ) KM/NIH mouse models were established, the mice were abdominally intravenously injected with MSCs, miR-223 inhibitor ( miR-223 in ) transfected MSCs or hypoxia-pretreated MSCs ( htMSCs ). Kidney tissues were harvested at 24 h after injection. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in blood samples were measured with ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ) and cysteine protease protein-3 ( caspase-3 ) in kidney tissue was detected by qRT-PCR. c Representative images from Western blot assays for apoptosis-related indicators. a P
Figure Legend Snippet: Knockdown of miR-223 blocked the protective effect of mesenchymal stem cells ( MSCs ) on renal tubular epithelial cells ( RTECs ). After the kidney ischemia/reperfusion ( IR ) KM/NIH mouse models were established, the mice were abdominally intravenously injected with MSCs, miR-223 inhibitor ( miR-223 in ) transfected MSCs or hypoxia-pretreated MSCs ( htMSCs ). Kidney tissues were harvested at 24 h after injection. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in blood samples were measured with ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ) and cysteine protease protein-3 ( caspase-3 ) in kidney tissue was detected by qRT-PCR. c Representative images from Western blot assays for apoptosis-related indicators. a P

Techniques Used: Mouse Assay, Injection, Transfection, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot

Expression of growth factors, chemokines, and antiapoptosis factors was suppressed by coculture with mesenchymal stem cells ( MSCs ) when miR-223 was inhibited; however, expression of proapoptosis factors was enhanced. All renal tubular epithelial cells ( RTECs ) were treated with hypoxia/reoxygenation ( HR ) stimulation. The MSCs had been transfected in advance with a miR-223 inhibitor ( miR-223 in ). The RTECs and MSCs were then cocultured in a double-chamber. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in coculture supernatants were measured with ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ), and cysteine protease protein-3 ( caspase-3 ) in RTECs was detected by qRT-PCR. c Representative images of Western blot assays for apoptosis-related indicators. a P
Figure Legend Snippet: Expression of growth factors, chemokines, and antiapoptosis factors was suppressed by coculture with mesenchymal stem cells ( MSCs ) when miR-223 was inhibited; however, expression of proapoptosis factors was enhanced. All renal tubular epithelial cells ( RTECs ) were treated with hypoxia/reoxygenation ( HR ) stimulation. The MSCs had been transfected in advance with a miR-223 inhibitor ( miR-223 in ). The RTECs and MSCs were then cocultured in a double-chamber. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in coculture supernatants were measured with ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ), and cysteine protease protein-3 ( caspase-3 ) in RTECs was detected by qRT-PCR. c Representative images of Western blot assays for apoptosis-related indicators. a P

Techniques Used: Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

Expression of growth factors, chemokines, and antiapoptosis factors was enhanced by coculture with mesenchymal stem cells ( MSCs ), while expression of proapoptosis factors was suppressed. Renal tubular epithelial cells ( RTECs ) were treated with hypoxia/reoxygenation ( HR ) stimulation and then cocultured with MSCs or hypoxia-pretreated MSCs ( htMSCs ) in a double-chamber. Cells were harvested at 48 h. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in coculture supernatants were measured by ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ), and cysteine protease protein-3 ( caspase-3 ) in RTECs was detected by qRT-PCR. c Representative images of Western blot assays for apoptosis-related indicators. a P
Figure Legend Snippet: Expression of growth factors, chemokines, and antiapoptosis factors was enhanced by coculture with mesenchymal stem cells ( MSCs ), while expression of proapoptosis factors was suppressed. Renal tubular epithelial cells ( RTECs ) were treated with hypoxia/reoxygenation ( HR ) stimulation and then cocultured with MSCs or hypoxia-pretreated MSCs ( htMSCs ) in a double-chamber. Cells were harvested at 48 h. a The concentrations of hepatocyte growth factor ( HGF ), insulin-like growth factor-1 ( IGF-1 ), transforming growth factor beta ( TGF-β ), and vascular endothelial growth factor ( VEGF ) in coculture supernatants were measured by ELISA. b Expression of B-cell lymphoma-2 ( Bcl-2 ), B-cell lymphoma-XL ( Bcl-XL ), cysteine protease protein-1 ( caspase-1 ), and cysteine protease protein-3 ( caspase-3 ) in RTECs was detected by qRT-PCR. c Representative images of Western blot assays for apoptosis-related indicators. a P

Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot

34) Product Images from "Matrix Metalloproteinase-2 Impairs Homing of Intracoronary Delivered Mesenchymal Stem Cells in a Porcine Reperfused Myocardial Infarction: Comparison With Intramyocardial Cell Delivery"

Article Title: Matrix Metalloproteinase-2 Impairs Homing of Intracoronary Delivered Mesenchymal Stem Cells in a Porcine Reperfused Myocardial Infarction: Comparison With Intramyocardial Cell Delivery

Journal: Frontiers in Bioengineering and Biotechnology

doi: 10.3389/fbioe.2018.00035

Expression of homing and angiogenic signals of the myocardium 7 days after cardiac transfer of green fluorescent protein (GFP)-Luc-mesenchymal stem cell (MSCs). Fluorescent immunohistochemistry of the bioluminescence positive myocardial areas 7 days after intramyocardial [left panel (A,C,E,G) ] or intracoronary [right panel (B,D,F,H) ] GFP-Luc-MSCs delivery shows increased expression of homing signals cadherin (A,B) , and angiogenic factors fibroblast growth factor 2 (FGF2) (C,D) and vascular endothelial growth factor (VEGF) (E,F) in group IM. Infarct area border zone (G,H) exhibited higher number of myocardial cells and higher level of VEGF expression in group IM (G) . Hoechst staining of the nuclei, 40× magnification. Expression of homing signals cadherin (I) , stromal-derived factor-1alpha (J) , and angiogenic factors FGF2 (K) and VEGF (L) .
Figure Legend Snippet: Expression of homing and angiogenic signals of the myocardium 7 days after cardiac transfer of green fluorescent protein (GFP)-Luc-mesenchymal stem cell (MSCs). Fluorescent immunohistochemistry of the bioluminescence positive myocardial areas 7 days after intramyocardial [left panel (A,C,E,G) ] or intracoronary [right panel (B,D,F,H) ] GFP-Luc-MSCs delivery shows increased expression of homing signals cadherin (A,B) , and angiogenic factors fibroblast growth factor 2 (FGF2) (C,D) and vascular endothelial growth factor (VEGF) (E,F) in group IM. Infarct area border zone (G,H) exhibited higher number of myocardial cells and higher level of VEGF expression in group IM (G) . Hoechst staining of the nuclei, 40× magnification. Expression of homing signals cadherin (I) , stromal-derived factor-1alpha (J) , and angiogenic factors FGF2 (K) and VEGF (L) .

Techniques Used: Expressing, Immunohistochemistry, Staining, Derivative Assay

35) Product Images from "Tiaogeng Yijing decoction improves the pregnancy outcomes of patients with poor ovarian response undergoing in vitro fertilization-embryo transfer"

Article Title: Tiaogeng Yijing decoction improves the pregnancy outcomes of patients with poor ovarian response undergoing in vitro fertilization-embryo transfer

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2017.4948

Tiaogeng Yijing increases the follicular fluid levels of GDF-9, TGF-β1 and VEGF in patients with POR
Figure Legend Snippet: Tiaogeng Yijing increases the follicular fluid levels of GDF-9, TGF-β1 and VEGF in patients with POR

Techniques Used:

Tiaogeng Yijing increases endometrial levels of integrin αVβ3, TGF-β1 and VEGF in patients with poor ovarian response. Endometrial protein expression levels of integrin αVβ3, TGF-β1, LIF, G-CSF and VEGF
Figure Legend Snippet: Tiaogeng Yijing increases endometrial levels of integrin αVβ3, TGF-β1 and VEGF in patients with poor ovarian response. Endometrial protein expression levels of integrin αVβ3, TGF-β1, LIF, G-CSF and VEGF

Techniques Used: Expressing

36) Product Images from "The Effect of Age on the Regenerative Potential of Human Eyelid Adipose-Derived Stem Cells"

Article Title: The Effect of Age on the Regenerative Potential of Human Eyelid Adipose-Derived Stem Cells

Journal: Stem Cells International

doi: 10.1155/2018/5654917

ELISA analysis of secreted proteins in HEASCs-CM. The graph showed that the expression of TGF- β 1 and cFn was decreased with advanced age; however, the expression of VEGF demonstrated an age-related increase in donor age groups. Data was represented as mean ± SD, ∗ P
Figure Legend Snippet: ELISA analysis of secreted proteins in HEASCs-CM. The graph showed that the expression of TGF- β 1 and cFn was decreased with advanced age; however, the expression of VEGF demonstrated an age-related increase in donor age groups. Data was represented as mean ± SD, ∗ P

Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing

37) Product Images from "Fibroblast activation protein in osteosarcoma cells promotes angiogenesis via AKT and ERK signaling pathways"

Article Title: Fibroblast activation protein in osteosarcoma cells promotes angiogenesis via AKT and ERK signaling pathways

Journal: Oncology Letters

doi: 10.3892/ol.2018.8027

FAP expression promotes VEGF-A expression in osteosarcoma cells. (A) Subsequent to the overexpression of FAP in MG63 cells, VEGF-A mRNA expression was upregulated, as determined with RT-PCR, which was (B) quantified by densitometry. (C) In addition, VEGF-A protein expression was upregulated, which was also (D) quantified. (E) Following the silencing of FAP expression in U2-OS cells, the expression of VEGF-A was inhibited at the mRNA level, as determined with RT-PCR, which was (F) quantified, and at (G) the protein level, which was (H) quantified. (I) The increase in VEGF-A subsequent to FAP overexpression and (J) the decrease in VEGF-A subsequent to FAP silencing were also confirmed by ELISA. *P
Figure Legend Snippet: FAP expression promotes VEGF-A expression in osteosarcoma cells. (A) Subsequent to the overexpression of FAP in MG63 cells, VEGF-A mRNA expression was upregulated, as determined with RT-PCR, which was (B) quantified by densitometry. (C) In addition, VEGF-A protein expression was upregulated, which was also (D) quantified. (E) Following the silencing of FAP expression in U2-OS cells, the expression of VEGF-A was inhibited at the mRNA level, as determined with RT-PCR, which was (F) quantified, and at (G) the protein level, which was (H) quantified. (I) The increase in VEGF-A subsequent to FAP overexpression and (J) the decrease in VEGF-A subsequent to FAP silencing were also confirmed by ELISA. *P

Techniques Used: Expressing, Over Expression, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

38) Product Images from "Association of gene polymorphism with serum levels of inflammatory and angiogenic factors in Pakistani patients with age-related macular degeneration"

Article Title: Association of gene polymorphism with serum levels of inflammatory and angiogenic factors in Pakistani patients with age-related macular degeneration

Journal: Molecular Vision

doi:

The box-and-whisker plot represents the mean and minimum to maximum range of IL-6, IL-8, VEGF and CRP levels in transformed data. Serum levels were measured in 100 control subjects and 90 AMD patients. Among AMD patients 63 had wet type and 27 had dry type AMD. The serum levels of ( A ) IL-6, ( B ) IL-8, ( C ) VEGF and (D) CRP were significantly elevated in AMD patients compared to control subjects (p
Figure Legend Snippet: The box-and-whisker plot represents the mean and minimum to maximum range of IL-6, IL-8, VEGF and CRP levels in transformed data. Serum levels were measured in 100 control subjects and 90 AMD patients. Among AMD patients 63 had wet type and 27 had dry type AMD. The serum levels of ( A ) IL-6, ( B ) IL-8, ( C ) VEGF and (D) CRP were significantly elevated in AMD patients compared to control subjects (p

Techniques Used: Whisker Assay, Transformation Assay

39) Product Images from "Transgenic overexpression of VEGF-C induces weight gain and insulin resistance in mice"

Article Title: Transgenic overexpression of VEGF-C induces weight gain and insulin resistance in mice

Journal: Scientific Reports

doi: 10.1038/srep31566

Reduced insulin sensitivity in K14-VEGF-C mice. ( a ) Elevated fasting blood glucose and insulin levels in K14-VEGF-C mice, leading to higher homeostatic model assessment of insulin resistance (HOMA-IR) indices. ( b ) Intraperitoneal glucose tolerance test ( n = 4–6 per group, mean ± SEM, *** p
Figure Legend Snippet: Reduced insulin sensitivity in K14-VEGF-C mice. ( a ) Elevated fasting blood glucose and insulin levels in K14-VEGF-C mice, leading to higher homeostatic model assessment of insulin resistance (HOMA-IR) indices. ( b ) Intraperitoneal glucose tolerance test ( n = 4–6 per group, mean ± SEM, *** p

Techniques Used: Mouse Assay

1 Increased weight gain and adipose tissue accumulation in K14-VEGF-C mice. ( a ) ELISA analysis revealed increased murine VEGF-C levels in SWAT of obese mice that were kept under HFD for 20 weeks. ( b ) Immunofluorescence analysis of tail skin showing enlarged lymphatic vessels (podoplanin, Pdpn, green) in K14-VEGF-C mice, while whole-mount immunostains of intestinal villi showed no differences in lacteal (LYVE-1, green) or blood vessel (CD31, red) structure between WT and K14-VEGF-C mice. ( c ) K14-VEGF-C mice gained more weight than WT littermates ( n = 8 per group, mean ± SEM is shown, * p
Figure Legend Snippet: 1 Increased weight gain and adipose tissue accumulation in K14-VEGF-C mice. ( a ) ELISA analysis revealed increased murine VEGF-C levels in SWAT of obese mice that were kept under HFD for 20 weeks. ( b ) Immunofluorescence analysis of tail skin showing enlarged lymphatic vessels (podoplanin, Pdpn, green) in K14-VEGF-C mice, while whole-mount immunostains of intestinal villi showed no differences in lacteal (LYVE-1, green) or blood vessel (CD31, red) structure between WT and K14-VEGF-C mice. ( c ) K14-VEGF-C mice gained more weight than WT littermates ( n = 8 per group, mean ± SEM is shown, * p

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Energy balance parameters of K14-VEGF-C mice are similar to WT controls. ( a ) Body weight, lean mass and fat mass, ( b ) Lean mass % and adiposity %. ( c ) Energy expenditure (normalized to body weight and analyzed with ANCOVA). ( d ) Activity, ( e ) food consumption, ( f ) VO 2 , ( g ) VCO 2 and (H) respiratory exchange rate (RER) were analyzed with indirect calorimetry using the Promethion system ( n = 6 per group). * p
Figure Legend Snippet: Energy balance parameters of K14-VEGF-C mice are similar to WT controls. ( a ) Body weight, lean mass and fat mass, ( b ) Lean mass % and adiposity %. ( c ) Energy expenditure (normalized to body weight and analyzed with ANCOVA). ( d ) Activity, ( e ) food consumption, ( f ) VO 2 , ( g ) VCO 2 and (H) respiratory exchange rate (RER) were analyzed with indirect calorimetry using the Promethion system ( n = 6 per group). * p

Techniques Used: Mouse Assay, Activity Assay

K14-VEGF-C mice show increased insulin resistance under HFD. ( a ) Increased body weight and ( b ) SWAT %, while EWAT % remained unaltered in K14-VEGF-C mice under HFD. ( c ) Fasting blood glucose was significantly elevated in K14-VEGF-C mice. ( d ) Insulin tolerance test ( n = 5 per group, mean ± SEM is shown, * p
Figure Legend Snippet: K14-VEGF-C mice show increased insulin resistance under HFD. ( a ) Increased body weight and ( b ) SWAT %, while EWAT % remained unaltered in K14-VEGF-C mice under HFD. ( c ) Fasting blood glucose was significantly elevated in K14-VEGF-C mice. ( d ) Insulin tolerance test ( n = 5 per group, mean ± SEM is shown, * p

Techniques Used: Mouse Assay

SWAT macrophages of K14-VEGF-C mice show enhanced pro-inflammatory characteristics before the onset of increased adiposity. ( a ) Data from 22-week-old mice, ( b–d ) data from 14-week-old mice; prior to the onset of weight gain. ( a ) Increased percentage of CD11b+ cells in SWAT stromal vascular fraction, with a significant elevation of M1/M2 macrophage marker ratios in K14-VEGF-C mice. ( b ) Gene expression analysis of M2 (CD163, CD206) and M1 (CD11c, TNF-α, IL6) markers showed a boosted M1 phenotype in isolated K14-VEGF-C SWAT macrophages from 14-week-old mice ( n = 3–4). ( c ) Increased BODIPY fluorescence signal revealed by enhanced OVA processing of K14-VEGF-C SWAT macrophages; histograms show the distribution of BODIPY signal intensity and the corresponding quantification per mouse ( n = 3 mice per group, scale bar = 100 μm). ( d ) Conditioned media from SWAT macrophages of K14-VEGF-C, but not WT mice, significantly reduced the differentiation of 3T3-L1 cells in vitro ( n = 4 mice per group, scale bar = 200 μm). ** p
Figure Legend Snippet: SWAT macrophages of K14-VEGF-C mice show enhanced pro-inflammatory characteristics before the onset of increased adiposity. ( a ) Data from 22-week-old mice, ( b–d ) data from 14-week-old mice; prior to the onset of weight gain. ( a ) Increased percentage of CD11b+ cells in SWAT stromal vascular fraction, with a significant elevation of M1/M2 macrophage marker ratios in K14-VEGF-C mice. ( b ) Gene expression analysis of M2 (CD163, CD206) and M1 (CD11c, TNF-α, IL6) markers showed a boosted M1 phenotype in isolated K14-VEGF-C SWAT macrophages from 14-week-old mice ( n = 3–4). ( c ) Increased BODIPY fluorescence signal revealed by enhanced OVA processing of K14-VEGF-C SWAT macrophages; histograms show the distribution of BODIPY signal intensity and the corresponding quantification per mouse ( n = 3 mice per group, scale bar = 100 μm). ( d ) Conditioned media from SWAT macrophages of K14-VEGF-C, but not WT mice, significantly reduced the differentiation of 3T3-L1 cells in vitro ( n = 4 mice per group, scale bar = 200 μm). ** p

Techniques Used: Mouse Assay, Marker, Expressing, Isolation, Fluorescence, In Vitro

40) Product Images from "Unexpected contribution of lymphatic vessels to promotion of distant metastatic tumor spread"

Article Title: Unexpected contribution of lymphatic vessels to promotion of distant metastatic tumor spread

Journal: Science Advances

doi: 10.1126/sciadv.aat4758

Characterization of CCSP-rtTA × tet-O–VEGF-C mice after doxycycline treatment for 2 weeks. ( A ) VEGF-C mRNA expression in the lung (normalized to RPLP0). The average expression level in WT × tet-O–VEGF-C mice was set to 1 ( n = 6). ( B ) VEGF-C protein levels in the lung detected by enzyme-linked immunosorbent assay (ELISA) ( n = 6). ( C ) Quantification of the VEGFR3 + LV area ( n = 4). ( D ) Representative images of VEGFR3 staining in the lung. Scale bars, 1 mm. ( E ) Representative high-magnification images of different regions in the lung stained for VEGFR3. Scale bars, 200 μm. PA, pulmonary artery; PV, pulmonary vein. ( F ) Representative images of MECA-32 staining in the lung. Scale bars, 50 μm. ( G ) Quantification of MECA-32 + blood vessel area in the lung ( n = 5).
Figure Legend Snippet: Characterization of CCSP-rtTA × tet-O–VEGF-C mice after doxycycline treatment for 2 weeks. ( A ) VEGF-C mRNA expression in the lung (normalized to RPLP0). The average expression level in WT × tet-O–VEGF-C mice was set to 1 ( n = 6). ( B ) VEGF-C protein levels in the lung detected by enzyme-linked immunosorbent assay (ELISA) ( n = 6). ( C ) Quantification of the VEGFR3 + LV area ( n = 4). ( D ) Representative images of VEGFR3 staining in the lung. Scale bars, 1 mm. ( E ) Representative high-magnification images of different regions in the lung stained for VEGFR3. Scale bars, 200 μm. PA, pulmonary artery; PV, pulmonary vein. ( F ) Representative images of MECA-32 staining in the lung. Scale bars, 50 μm. ( G ) Quantification of MECA-32 + blood vessel area in the lung ( n = 5).

Techniques Used: Mouse Assay, Expressing, Enzyme-linked Immunosorbent Assay, Staining

Increased metastasis in CCSP-rtTA × tet-O–VEGF-C mice after tail vein injection of B16F10 melanoma cells. ( A ) Schematic of the B16F10 tumor study. Dox, doxycycline; i.v., intravenous. ( B ) Representative images of lung sections with metastatic nodules 14 days after tumor cell injection (indicated by staining for RFP from B16F10-luc2-tdTomato cells). Scale bars, 1 mm. ( C ) Quantification of tdTomato + area in the lung 14 days after tumor cell injection ( n = 10). ( D ) Quantification of tdTomato + cells per lung 24 hours after intravenous injection of 1 × 10 6 B16F10-luc2-tdTomato cells. ( E ) Representative images of lymphatic invasion (white arrows) with orthogonal projection. Scale bars, 100 μm. Fisher’s exact test was performed. ( F ) Quantification of the lung-draining lymph node weight ( n = 10). ( G ) Number of mice with different numbers of distant organs with metastasis (zero, one, two, or three organs affected) and example images of metastases (yellow arrows) in the liver and intestine. Data were pooled from two rounds of studies.
Figure Legend Snippet: Increased metastasis in CCSP-rtTA × tet-O–VEGF-C mice after tail vein injection of B16F10 melanoma cells. ( A ) Schematic of the B16F10 tumor study. Dox, doxycycline; i.v., intravenous. ( B ) Representative images of lung sections with metastatic nodules 14 days after tumor cell injection (indicated by staining for RFP from B16F10-luc2-tdTomato cells). Scale bars, 1 mm. ( C ) Quantification of tdTomato + area in the lung 14 days after tumor cell injection ( n = 10). ( D ) Quantification of tdTomato + cells per lung 24 hours after intravenous injection of 1 × 10 6 B16F10-luc2-tdTomato cells. ( E ) Representative images of lymphatic invasion (white arrows) with orthogonal projection. Scale bars, 100 μm. Fisher’s exact test was performed. ( F ) Quantification of the lung-draining lymph node weight ( n = 10). ( G ) Number of mice with different numbers of distant organs with metastasis (zero, one, two, or three organs affected) and example images of metastases (yellow arrows) in the liver and intestine. Data were pooled from two rounds of studies.

Techniques Used: Mouse Assay, Injection, Staining

Increased metastasis in CCSP-rtTA × tet-O–VEGF-C mice in the 4T1 spontaneous breast cancer metastasis model. ( A ) Schematic of the 4T1 tumor study. PR, postremoval; s.c., subcutaneous. ( B ) Representative in vivo bioluminescence images of metastases on day 21 after primary tumor removal. ( C ) Quantification of bioluminescence signal over the whole body on days PR7, PR14, and PR21 by IVIS imaging. For data from days PR7 and PR14, WT × VEGFC, n = 8, and CCSP × VEGFC, n = 11. Four mice reached the euthanization criteria before the end time point; thus, for data from day PR 21, WT × VEGFC, n = 7, and CCSP × VEGFC, n = 8. ns, not significant. ( D ) Number of mice with different numbers of distant organs with metastasis (zero, one, two, or three organs affected). Data were pooled from three rounds of studies. Only mice with detectable metastases (by ex vivo IVIS imaging) in the lungs were quantified.
Figure Legend Snippet: Increased metastasis in CCSP-rtTA × tet-O–VEGF-C mice in the 4T1 spontaneous breast cancer metastasis model. ( A ) Schematic of the 4T1 tumor study. PR, postremoval; s.c., subcutaneous. ( B ) Representative in vivo bioluminescence images of metastases on day 21 after primary tumor removal. ( C ) Quantification of bioluminescence signal over the whole body on days PR7, PR14, and PR21 by IVIS imaging. For data from days PR7 and PR14, WT × VEGFC, n = 8, and CCSP × VEGFC, n = 11. Four mice reached the euthanization criteria before the end time point; thus, for data from day PR 21, WT × VEGFC, n = 7, and CCSP × VEGFC, n = 8. ns, not significant. ( D ) Number of mice with different numbers of distant organs with metastasis (zero, one, two, or three organs affected). Data were pooled from three rounds of studies. Only mice with detectable metastases (by ex vivo IVIS imaging) in the lungs were quantified.

Techniques Used: Mouse Assay, In Vivo, Imaging, Ex Vivo

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Enzyme-linked Immunosorbent Assay:

Article Title: The Evaluation of the Relationship between sTREM-1, VEGF-B, and VEGF Gene Expression Levels with Disease Activity of Behçet's Patients
Article Snippet: .. Centrifuged serum samples were stored at −80°C until the working day. sTREM-1, VEGF-B, and VEGF gene expression levels were studied from the patients and from the healthy volunteers. sTREM-1 is assessed with Cusabio Biotech Co. limited kit (Wuhan, China) by using the ELISA method from the serum. sTREM-1 interassay CV values are 12.6% while the intraassay CV values are 10.7%. ..

Article Title: Fibroblast activation protein in osteosarcoma cells promotes angiogenesis via AKT and ERK signaling pathways
Article Snippet: .. VEGF-A in CM was detected using PGRN ELISA kit (cat. no. CSB-E11718h; CUSABIO Life Science, Wuhan, China) and VEGF-A ELISA Kit (CUSABIO Life Science) according to the manufacturer's instructions. ..

Article Title: The Effect of Age on the Regenerative Potential of Human Eyelid Adipose-Derived Stem Cells
Article Snippet: .. ELISA was performed using a human platelet-derived growth factor-BB (PDGF-BB), human transforming growth factor β 1 (TGF-β 1), human vascular endothelial cell growth factor (VEGF), human cellular fibronectin (cFn), human collagen type I (Col I), human interleukin 8 (IL-8) ELISA kit (CUSABIO, China), and their protocols. .. As a reference for quantification, a standard curve was established by a serial dilution for PDGF-BB protein (0.156 ng/ml–10 ng/ml), TGF-β 1 protein (0.78 ng/ml–50 ng/ml), VEGF protein (31.25 pg/ml–2000 pg/ml), cFn protein (12.5 ng/ml–800 ng/ml), Col-1 protein (1.56 ng/ml–100 ng/ml), and IL-8 protein (31.25 pg/ml–2000 pg/ml).

Article Title: Plastic roles of pericytes in the blood–retinal barrier
Article Snippet: .. To measure the intravitreal levels of soluble Tie1 or VEGF-A, after 40-fold dilution of vitreous sample, mouse Tie1 ELISA kit (CUSABIO, CSB-E07391m) or mouse VEGF-A ELISA kit (R & D, MMV00) were used according to the manufacturer's instruction and measured using a Spectra MAX340 plate reader (Molecular Devices). .. Chromatin immunoprecipitation-qPCR ∼5 × 106 HUVECs were harvested and fixed with 11% formaldehyde solution and quenched by 2.5 M glycine.

Article Title: Metformin prevents hormonal and metabolic disturbances and 1,2-dimethylhydrazine-induced colon carcinogenesis in non-diabetic rats
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Article Title: Therapeutic Lymphangiogenesis With Implantation of Adipose-Derived Regenerative Cells
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Article Title: Ethyl-p-methoxycinnamate isolated from kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-?, and angiogenesis by blocking endothelial functions
Article Snippet: .. Human IL-1, human TNF-α, human nitric oxide (NO), human vascular endothelial growth factor (VEGF), rat IL-1, and rat TNF-α ELISA kits were purchased from Cusabio, China. .. RPMI 1640, human umbilical vein endothelial cells (HUVEC), and endothelial cell medium (ECM) supplied with endothelial cell growth supplements (ECGS) were obtained from ScienCell, USA.

Article Title: Interactions between fibroblastic reticular cells and B cells promote mesenteric lymph node lymphangiogenesis
Article Snippet: .. VEGF-A levels in the culture supernatant or in tissue homogenates were determined using mouse VEGF quantikine ELISA kit (R & D system, Cat No. MMV00), where as VEGF-C levels were determined using mouse VEGF-C ELISA kit (CUSABIO, CSB-E07361m) according to the manufacturer’s instructions. .. In another set of experiments, mice were sacrificed at indicated time point post infection and intracellular VEGFs expressing B cells were measured as described previously with out any further manipulations .

Expressing:

Article Title: The Evaluation of the Relationship between sTREM-1, VEGF-B, and VEGF Gene Expression Levels with Disease Activity of Behçet's Patients
Article Snippet: .. Centrifuged serum samples were stored at −80°C until the working day. sTREM-1, VEGF-B, and VEGF gene expression levels were studied from the patients and from the healthy volunteers. sTREM-1 is assessed with Cusabio Biotech Co. limited kit (Wuhan, China) by using the ELISA method from the serum. sTREM-1 interassay CV values are 12.6% while the intraassay CV values are 10.7%. ..

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    Cusabio vegf a
    Additional <t>VEGF-A</t> is required to recapitulate the phenotypes of DR. ( a ) Diagram depicting the experiment schedule for selective loss of pericytes in retinal vessels using adult DTA iΔPC mice, intra-vitreal administration of VEGF-A (1 μg) into one eye and BSA (1 μg) into the contralateral eye, and analyses at 4 days after the administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, PDGFRβ + pericyte coverage, expressions of Ang2 and VEGFR2 in CD31 + vessels, and dextran (70 kDa) leakage in retinas of WT and DTA iΔPC mice that were intra-vitreally treated with BSA (B) or VEGF-A (VA). Each group, n =6. Error bars represent mean±s.d. * P
    Vegf A, supplied by Cusabio, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cusabio mouse vegf c kit
    Vegfr3 -deleted LECs induce proliferation of neighboring VEGFR3 + cells in a cell-contact-dependent manner. a <t>VEGF-C</t> concentration measured by <t>ELISA</t> in the ear skin at indicated stages. Bars represent mean ( n = 2–8 mice, as indicated) ± s.d. b Experimental set up for co-cultures of WT and VEGFR3 − (KO) primary LECs from Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice (left) and immunofluorescence showing VEGFR3 depletion in Cre − targeted (GFP + ) LECs (right, arrows). c Left: proliferating WT cells were frequently associated with KO cells. Right: quantification of the proportion of EdU + WT cells that were in direct contact with KO cells (mean ( n = 6 biological replicates) ± s.e.m). d Assessment of cell proliferation in co-cultures of primary human LECs treated with control (siCTRL) or VEGFR3 (siVEGFR3) siRNA. Cell tracker-labeled siCTRL cells (purple) were mixed with unlabeled siVEGFR3 cells in 40:60%, or they were cultured alone. e Quantification of proliferating EdU + cells in co-cultures of siCTRL and siVEGFR3 LECs mixed in different ratios. Data are normalized to the proliferation rate (0.8 ± 0.2%) in siCTRL alone and represent mean ( n = 5 biological replicates) ± s.e.m. f Predicted and observed proportions of siCTRL LECs (all vs. EdU + ) that are in direct contact with siVEGFR3 LECs in 80:20% siCTRL:siVEGFR3 co-cultures (mean ( n = 5 biological replicates) ± s.e.m). Predicted proportion was calculated based on random distribution and n = 5 (5.2 ± 0.2, quantified from n = 172 LECs stained for VE-cadherin) neighbors. g Quantification of proliferating EdU + cells in siCTRL LECs cultured alone (−), or co-cultured without direct cell–cell contacts with siVEGFR3 cells seeded on Transwell inserts (+). Data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m. * P
    Mouse Vegf C Kit, supplied by Cusabio, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BAFF and IL-4 stimulation of B cells promotes lymphotoxin and VEGFs production. a – c mLN B cells were cultured with anti-IgM with or without IL-4±BAFF and the culture supernatant collected at a 72 h or b 18 h post stimulation for quantitation of VEGF-A and <t>VEGF-C</t> by <t>ELISA.</t> c Splenic B cells were stimulated as in ( a ) and the culture supernatant collected at 72 h for quantitation of VEGF-A and VEGF-C by ELISA. Data represent mean ± SEM, and are representative of pooled data from three independent experiments a or are representative of two independent experiments b , c . d Naive mLN B cells were cultured with or without IL-4±BAFF for 18 h then B-cell lymphotoxin expression determined by staining with LTβR-Fc. Data represent mean ± SEM and representative of three independent experiments. e mLN cryosections from mixed BMC-lacking lymphotoxin on B or T cells and infected with Hp for 21 days were stained for BAFF + ( red ) and FRCs (ER-TR7 + Laminin + ). Scale bar = 100 μm. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P
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    Additional VEGF-A is required to recapitulate the phenotypes of DR. ( a ) Diagram depicting the experiment schedule for selective loss of pericytes in retinal vessels using adult DTA iΔPC mice, intra-vitreal administration of VEGF-A (1 μg) into one eye and BSA (1 μg) into the contralateral eye, and analyses at 4 days after the administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, PDGFRβ + pericyte coverage, expressions of Ang2 and VEGFR2 in CD31 + vessels, and dextran (70 kDa) leakage in retinas of WT and DTA iΔPC mice that were intra-vitreally treated with BSA (B) or VEGF-A (VA). Each group, n =6. Error bars represent mean±s.d. * P

    Journal: Nature Communications

    Article Title: Plastic roles of pericytes in the blood–retinal barrier

    doi: 10.1038/ncomms15296

    Figure Lengend Snippet: Additional VEGF-A is required to recapitulate the phenotypes of DR. ( a ) Diagram depicting the experiment schedule for selective loss of pericytes in retinal vessels using adult DTA iΔPC mice, intra-vitreal administration of VEGF-A (1 μg) into one eye and BSA (1 μg) into the contralateral eye, and analyses at 4 days after the administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, PDGFRβ + pericyte coverage, expressions of Ang2 and VEGFR2 in CD31 + vessels, and dextran (70 kDa) leakage in retinas of WT and DTA iΔPC mice that were intra-vitreally treated with BSA (B) or VEGF-A (VA). Each group, n =6. Error bars represent mean±s.d. * P

    Article Snippet: To measure the intravitreal levels of soluble Tie1 or VEGF-A, after 40-fold dilution of vitreous sample, mouse Tie1 ELISA kit (CUSABIO, CSB-E07391m) or mouse VEGF-A ELISA kit (R & D, MMV00) were used according to the manufacturer's instruction and measured using a Spectra MAX340 plate reader (Molecular Devices).

    Techniques: Mouse Assay

    Tie2 activation lessens BRB disintegration in pericyte-free adult retina. ( a ) Diagram depicting the experiment schedule for selective depletion of pericytes around retinal vessels by tamoxifen, intra-vitreal administration of VEGF-A (1 μg), and daily intra-peritoneal administration of ABTTA (20 mg kg −1 ) for 4 days in adult DTA iΔPC mice. Analyses were performed 4 days after VEGF-A administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, dextran (70 kDa) leakage, and F4/80 + macrophage infiltration in VA-DTA iΔPC mice treated with Fc (Fc; n =6) or ABTAA (AB; n =6). Boxed regions (dotted-lines) are magnified and presented in panels in the corner. Error bars represent mean±s.d. ** P

    Journal: Nature Communications

    Article Title: Plastic roles of pericytes in the blood–retinal barrier

    doi: 10.1038/ncomms15296

    Figure Lengend Snippet: Tie2 activation lessens BRB disintegration in pericyte-free adult retina. ( a ) Diagram depicting the experiment schedule for selective depletion of pericytes around retinal vessels by tamoxifen, intra-vitreal administration of VEGF-A (1 μg), and daily intra-peritoneal administration of ABTTA (20 mg kg −1 ) for 4 days in adult DTA iΔPC mice. Analyses were performed 4 days after VEGF-A administration. ( b , c ) Images and comparisons of inner surface of retinal cup, CD31 + vessels, dextran (70 kDa) leakage, and F4/80 + macrophage infiltration in VA-DTA iΔPC mice treated with Fc (Fc; n =6) or ABTAA (AB; n =6). Boxed regions (dotted-lines) are magnified and presented in panels in the corner. Error bars represent mean±s.d. ** P

    Article Snippet: To measure the intravitreal levels of soluble Tie1 or VEGF-A, after 40-fold dilution of vitreous sample, mouse Tie1 ELISA kit (CUSABIO, CSB-E07391m) or mouse VEGF-A ELISA kit (R & D, MMV00) were used according to the manufacturer's instruction and measured using a Spectra MAX340 plate reader (Molecular Devices).

    Techniques: Activation Assay, Mouse Assay

    FAP expression promotes VEGF-A expression in osteosarcoma cells. (A) Subsequent to the overexpression of FAP in MG63 cells, VEGF-A mRNA expression was upregulated, as determined with RT-PCR, which was (B) quantified by densitometry. (C) In addition, VEGF-A protein expression was upregulated, which was also (D) quantified. (E) Following the silencing of FAP expression in U2-OS cells, the expression of VEGF-A was inhibited at the mRNA level, as determined with RT-PCR, which was (F) quantified, and at (G) the protein level, which was (H) quantified. (I) The increase in VEGF-A subsequent to FAP overexpression and (J) the decrease in VEGF-A subsequent to FAP silencing were also confirmed by ELISA. *P

    Journal: Oncology Letters

    Article Title: Fibroblast activation protein in osteosarcoma cells promotes angiogenesis via AKT and ERK signaling pathways

    doi: 10.3892/ol.2018.8027

    Figure Lengend Snippet: FAP expression promotes VEGF-A expression in osteosarcoma cells. (A) Subsequent to the overexpression of FAP in MG63 cells, VEGF-A mRNA expression was upregulated, as determined with RT-PCR, which was (B) quantified by densitometry. (C) In addition, VEGF-A protein expression was upregulated, which was also (D) quantified. (E) Following the silencing of FAP expression in U2-OS cells, the expression of VEGF-A was inhibited at the mRNA level, as determined with RT-PCR, which was (F) quantified, and at (G) the protein level, which was (H) quantified. (I) The increase in VEGF-A subsequent to FAP overexpression and (J) the decrease in VEGF-A subsequent to FAP silencing were also confirmed by ELISA. *P

    Article Snippet: VEGF-A in CM was detected using PGRN ELISA kit (cat. no. CSB-E11718h; CUSABIO Life Science, Wuhan, China) and VEGF-A ELISA Kit (CUSABIO Life Science) according to the manufacturer's instructions.

    Techniques: Expressing, Over Expression, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    FAP and VEGF-A expression in osteosarcoma cells. FAP and VEGF-A expression were detected in MG63, U2-OS and HOS cell lines. (A) FAP and VEGF-A mRNA expression levels were determined with reverse transcription-polymerase chain reaction, which was (B) quantified by densitometry. (C) FAP and VEGF-A protein expression levels were determined with western blot analysis, which was (D) quantified by densitometry. FAP and VEGF-A in U2-OS cells was compared with the other two groups. *P

    Journal: Oncology Letters

    Article Title: Fibroblast activation protein in osteosarcoma cells promotes angiogenesis via AKT and ERK signaling pathways

    doi: 10.3892/ol.2018.8027

    Figure Lengend Snippet: FAP and VEGF-A expression in osteosarcoma cells. FAP and VEGF-A expression were detected in MG63, U2-OS and HOS cell lines. (A) FAP and VEGF-A mRNA expression levels were determined with reverse transcription-polymerase chain reaction, which was (B) quantified by densitometry. (C) FAP and VEGF-A protein expression levels were determined with western blot analysis, which was (D) quantified by densitometry. FAP and VEGF-A in U2-OS cells was compared with the other two groups. *P

    Article Snippet: VEGF-A in CM was detected using PGRN ELISA kit (cat. no. CSB-E11718h; CUSABIO Life Science, Wuhan, China) and VEGF-A ELISA Kit (CUSABIO Life Science) according to the manufacturer's instructions.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

    Vegfr3 -deleted LECs induce proliferation of neighboring VEGFR3 + cells in a cell-contact-dependent manner. a VEGF-C concentration measured by ELISA in the ear skin at indicated stages. Bars represent mean ( n = 2–8 mice, as indicated) ± s.d. b Experimental set up for co-cultures of WT and VEGFR3 − (KO) primary LECs from Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice (left) and immunofluorescence showing VEGFR3 depletion in Cre − targeted (GFP + ) LECs (right, arrows). c Left: proliferating WT cells were frequently associated with KO cells. Right: quantification of the proportion of EdU + WT cells that were in direct contact with KO cells (mean ( n = 6 biological replicates) ± s.e.m). d Assessment of cell proliferation in co-cultures of primary human LECs treated with control (siCTRL) or VEGFR3 (siVEGFR3) siRNA. Cell tracker-labeled siCTRL cells (purple) were mixed with unlabeled siVEGFR3 cells in 40:60%, or they were cultured alone. e Quantification of proliferating EdU + cells in co-cultures of siCTRL and siVEGFR3 LECs mixed in different ratios. Data are normalized to the proliferation rate (0.8 ± 0.2%) in siCTRL alone and represent mean ( n = 5 biological replicates) ± s.e.m. f Predicted and observed proportions of siCTRL LECs (all vs. EdU + ) that are in direct contact with siVEGFR3 LECs in 80:20% siCTRL:siVEGFR3 co-cultures (mean ( n = 5 biological replicates) ± s.e.m). Predicted proportion was calculated based on random distribution and n = 5 (5.2 ± 0.2, quantified from n = 172 LECs stained for VE-cadherin) neighbors. g Quantification of proliferating EdU + cells in siCTRL LECs cultured alone (−), or co-cultured without direct cell–cell contacts with siVEGFR3 cells seeded on Transwell inserts (+). Data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m. * P

    Journal: Nature Communications

    Article Title: Heterogeneity in VEGFR3 levels drives lymphatic vessel hyperplasia through cell-autonomous and non-cell-autonomous mechanisms

    doi: 10.1038/s41467-018-03692-0

    Figure Lengend Snippet: Vegfr3 -deleted LECs induce proliferation of neighboring VEGFR3 + cells in a cell-contact-dependent manner. a VEGF-C concentration measured by ELISA in the ear skin at indicated stages. Bars represent mean ( n = 2–8 mice, as indicated) ± s.d. b Experimental set up for co-cultures of WT and VEGFR3 − (KO) primary LECs from Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice (left) and immunofluorescence showing VEGFR3 depletion in Cre − targeted (GFP + ) LECs (right, arrows). c Left: proliferating WT cells were frequently associated with KO cells. Right: quantification of the proportion of EdU + WT cells that were in direct contact with KO cells (mean ( n = 6 biological replicates) ± s.e.m). d Assessment of cell proliferation in co-cultures of primary human LECs treated with control (siCTRL) or VEGFR3 (siVEGFR3) siRNA. Cell tracker-labeled siCTRL cells (purple) were mixed with unlabeled siVEGFR3 cells in 40:60%, or they were cultured alone. e Quantification of proliferating EdU + cells in co-cultures of siCTRL and siVEGFR3 LECs mixed in different ratios. Data are normalized to the proliferation rate (0.8 ± 0.2%) in siCTRL alone and represent mean ( n = 5 biological replicates) ± s.e.m. f Predicted and observed proportions of siCTRL LECs (all vs. EdU + ) that are in direct contact with siVEGFR3 LECs in 80:20% siCTRL:siVEGFR3 co-cultures (mean ( n = 5 biological replicates) ± s.e.m). Predicted proportion was calculated based on random distribution and n = 5 (5.2 ± 0.2, quantified from n = 172 LECs stained for VE-cadherin) neighbors. g Quantification of proliferating EdU + cells in siCTRL LECs cultured alone (−), or co-cultured without direct cell–cell contacts with siVEGFR3 cells seeded on Transwell inserts (+). Data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m. * P

    Article Snippet: Three micrograms of total protein was used for ELISA assay using mouse VEGF-C kit (CUSABIO).

    Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Mouse Assay, Immunofluorescence, Labeling, Cell Culture, Staining

    Global and mosaic loss of DLL4 promote LEC proliferation through inhibition of Notch signaling in neighboring cells. a , b DAPT induced effect on the proliferation of control (siCTRL) or VEGFR3 (siVEGFR3) siRNA-treated LECs. a Tile scan immunofluorescence images and ( b ) quantification of proliferating EdU + LECs after 3 h of incorporation (mean ( n = 3 biological replicates) ± s.e.m). c Western blot analysis of cell lysates from LECs treated with siCTRL or NOTCH1 siRNA (siNOTCH1) and ( d ) quantification of proliferating EdU + LECs cultured with or without VEGF-C (data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 3 biological replicates) ± s.e.m). e Western blot analysis of cell lysates from LECs treated with siCTRL or DLL4 siRNA (siDLL4) and ( f ) quantification of proliferating EdU + cells in co-cultures of siCTRL and siDLL4 LECs mixed in different ratios (data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m). g Model of cell-autonomous and non-cell-autonomous effects of VEGFR3 expression defining LEC responses during sprouting lymphangiogenesis. VEGF-C-VEGFR3 signaling positively regulates DLL4 expression leading to activation of Notch and cell cycle arrest in neighboring LECs. VEGFR3 deficiency (green cell) consequently leads to downregulation of DLL4 and inhibition of Notch in neighboring LECs, but only VEGFR3 expressing (red) cells respond by proliferation and sprouting. When VEGFR3 − cells are in excess, VEGFR3 + cells interact predominantly with LECs with low DLL4 and respond by proliferation (dashed line) * P

    Journal: Nature Communications

    Article Title: Heterogeneity in VEGFR3 levels drives lymphatic vessel hyperplasia through cell-autonomous and non-cell-autonomous mechanisms

    doi: 10.1038/s41467-018-03692-0

    Figure Lengend Snippet: Global and mosaic loss of DLL4 promote LEC proliferation through inhibition of Notch signaling in neighboring cells. a , b DAPT induced effect on the proliferation of control (siCTRL) or VEGFR3 (siVEGFR3) siRNA-treated LECs. a Tile scan immunofluorescence images and ( b ) quantification of proliferating EdU + LECs after 3 h of incorporation (mean ( n = 3 biological replicates) ± s.e.m). c Western blot analysis of cell lysates from LECs treated with siCTRL or NOTCH1 siRNA (siNOTCH1) and ( d ) quantification of proliferating EdU + LECs cultured with or without VEGF-C (data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 3 biological replicates) ± s.e.m). e Western blot analysis of cell lysates from LECs treated with siCTRL or DLL4 siRNA (siDLL4) and ( f ) quantification of proliferating EdU + cells in co-cultures of siCTRL and siDLL4 LECs mixed in different ratios (data are normalized to the proliferation rate in siCTRL alone and represent mean ( n = 6 biological replicates) ± s.e.m). g Model of cell-autonomous and non-cell-autonomous effects of VEGFR3 expression defining LEC responses during sprouting lymphangiogenesis. VEGF-C-VEGFR3 signaling positively regulates DLL4 expression leading to activation of Notch and cell cycle arrest in neighboring LECs. VEGFR3 deficiency (green cell) consequently leads to downregulation of DLL4 and inhibition of Notch in neighboring LECs, but only VEGFR3 expressing (red) cells respond by proliferation and sprouting. When VEGFR3 − cells are in excess, VEGFR3 + cells interact predominantly with LECs with low DLL4 and respond by proliferation (dashed line) * P

    Article Snippet: Three micrograms of total protein was used for ELISA assay using mouse VEGF-C kit (CUSABIO).

    Techniques: Inhibition, Immunofluorescence, Western Blot, Cell Culture, Expressing, Activation Assay

    Lymphatic vascular hyperplasia in Vegfr3 -deleted ears is driven by VEGF-C signaling. a Quantification of blunt-ended vessels (lymphatic capillaries) in 5 weeks old mice treated with Tamoxifen at P2, P4, and P6. Central region excludes 560 μm area (tip) from the edge of the ear. Bars represent mean ( n = 3–6 mice, as indicated) ± s.e.m. b Whole-mount immunofluorescence of Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 ear skin showing vessel interconnections (arrows) formed of non-targeted (GFP − ) LYVE1 + (left) and VEGFR3 + (right) LECs. c Whole-mount immunofluorescence of ear skin of 3 weeks old Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 and Cre-negative littermate mice treated with Tamoxifen at P2, P4, and P6, and inhibitors of VEGF-C signaling (the VEGF-C-trap VEGFR3-Ig, the VEGFR3 inhibitor MAZ51, or the VEGFR2 blocking antibody DC101). Quantification of ( d ) vessel area and ( e ) branch points. Bars represent mean ( n = 3 (untreated groups) or n = 4 (treated groups) mice) ± s.e.m. f – h Visualization by whole-mount immunofluorescence (arrows in f ) and quantification of blunt-ended vessels in the ( g ) entire and ( h ) central region of the ear in Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice treated with the VEGFR3 kinase inhibitor MAZ51. In g , h , bars represent mean ( n = 3 (untreated) or n = 4 (MAZ51, DC101) mice) ± s.e.m. * P

    Journal: Nature Communications

    Article Title: Heterogeneity in VEGFR3 levels drives lymphatic vessel hyperplasia through cell-autonomous and non-cell-autonomous mechanisms

    doi: 10.1038/s41467-018-03692-0

    Figure Lengend Snippet: Lymphatic vascular hyperplasia in Vegfr3 -deleted ears is driven by VEGF-C signaling. a Quantification of blunt-ended vessels (lymphatic capillaries) in 5 weeks old mice treated with Tamoxifen at P2, P4, and P6. Central region excludes 560 μm area (tip) from the edge of the ear. Bars represent mean ( n = 3–6 mice, as indicated) ± s.e.m. b Whole-mount immunofluorescence of Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 ear skin showing vessel interconnections (arrows) formed of non-targeted (GFP − ) LYVE1 + (left) and VEGFR3 + (right) LECs. c Whole-mount immunofluorescence of ear skin of 3 weeks old Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 and Cre-negative littermate mice treated with Tamoxifen at P2, P4, and P6, and inhibitors of VEGF-C signaling (the VEGF-C-trap VEGFR3-Ig, the VEGFR3 inhibitor MAZ51, or the VEGFR2 blocking antibody DC101). Quantification of ( d ) vessel area and ( e ) branch points. Bars represent mean ( n = 3 (untreated groups) or n = 4 (treated groups) mice) ± s.e.m. f – h Visualization by whole-mount immunofluorescence (arrows in f ) and quantification of blunt-ended vessels in the ( g ) entire and ( h ) central region of the ear in Vegfr3 flox/flox ; R26-mTmG;Prox1-CreER T2 mice treated with the VEGFR3 kinase inhibitor MAZ51. In g , h , bars represent mean ( n = 3 (untreated) or n = 4 (MAZ51, DC101) mice) ± s.e.m. * P

    Article Snippet: Three micrograms of total protein was used for ELISA assay using mouse VEGF-C kit (CUSABIO).

    Techniques: Mouse Assay, Immunofluorescence, Blocking Assay

    BAFF and IL-4 stimulation of B cells promotes lymphotoxin and VEGFs production. a – c mLN B cells were cultured with anti-IgM with or without IL-4±BAFF and the culture supernatant collected at a 72 h or b 18 h post stimulation for quantitation of VEGF-A and VEGF-C by ELISA. c Splenic B cells were stimulated as in ( a ) and the culture supernatant collected at 72 h for quantitation of VEGF-A and VEGF-C by ELISA. Data represent mean ± SEM, and are representative of pooled data from three independent experiments a or are representative of two independent experiments b , c . d Naive mLN B cells were cultured with or without IL-4±BAFF for 18 h then B-cell lymphotoxin expression determined by staining with LTβR-Fc. Data represent mean ± SEM and representative of three independent experiments. e mLN cryosections from mixed BMC-lacking lymphotoxin on B or T cells and infected with Hp for 21 days were stained for BAFF + ( red ) and FRCs (ER-TR7 + Laminin + ). Scale bar = 100 μm. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P

    Journal: Nature Communications

    Article Title: Interactions between fibroblastic reticular cells and B cells promote mesenteric lymph node lymphangiogenesis

    doi: 10.1038/s41467-017-00504-9

    Figure Lengend Snippet: BAFF and IL-4 stimulation of B cells promotes lymphotoxin and VEGFs production. a – c mLN B cells were cultured with anti-IgM with or without IL-4±BAFF and the culture supernatant collected at a 72 h or b 18 h post stimulation for quantitation of VEGF-A and VEGF-C by ELISA. c Splenic B cells were stimulated as in ( a ) and the culture supernatant collected at 72 h for quantitation of VEGF-A and VEGF-C by ELISA. Data represent mean ± SEM, and are representative of pooled data from three independent experiments a or are representative of two independent experiments b , c . d Naive mLN B cells were cultured with or without IL-4±BAFF for 18 h then B-cell lymphotoxin expression determined by staining with LTβR-Fc. Data represent mean ± SEM and representative of three independent experiments. e mLN cryosections from mixed BMC-lacking lymphotoxin on B or T cells and infected with Hp for 21 days were stained for BAFF + ( red ) and FRCs (ER-TR7 + Laminin + ). Scale bar = 100 μm. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P

    Article Snippet: VEGF-A levels in the culture supernatant or in tissue homogenates were determined using mouse VEGF quantikine ELISA kit (R & D system, Cat No. MMV00), where as VEGF-C levels were determined using mouse VEGF-C ELISA kit (CUSABIO, CSB-E07361m) according to the manufacturer’s instructions.

    Techniques: Cell Culture, Quantitation Assay, Enzyme-linked Immunosorbent Assay, Expressing, Staining, Infection

    In vivo BAFF inhibition attenuates helminth-induced lymphangiogenesis. a C57BL/6 J wild-type mice were treated with isotype control or anti-BAFF mAb (Sandy-2) and infected with Hp . The entire chain of the mLN was collected at 12 dpi and processed for flow cytometry or histological staining. b Total weight of mLN, c total cell count, d total number of B cells, e absolute number of lymphotoxin-expressing B cells, and f total FRCs (PDPN + MadCAM1 − CD31 − ) present within the mLN as determined using flow cytometry. g mLN serial cryosections showing lymphatic organization after treatment with isotype control or anti-BAFF mAbs in naive or at 12 dpi mice ( blue ; DAPI, grays ; LYVE-1 + LECs). Scale bar = 2000 μm. Images are from a single mouse and are representative of two independent experiments each including n ≥ 2 mice/group/time point. h VEGF-A and i VEGF-C in mLN tissue homogenates as determined by ELISA. j , k total LECs (PDPN + CD-31 + ) and proliferating LECs (PDPN + CD-31 + Ki-67 + ) in the mLN of naive and 12 dpi mice treated with isotype control or anti-BAFF mAbs were determined using flow cytometry. Data represent mean ± SEM and representative of two independent experiments with n = 3mice/group/time-point. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P

    Journal: Nature Communications

    Article Title: Interactions between fibroblastic reticular cells and B cells promote mesenteric lymph node lymphangiogenesis

    doi: 10.1038/s41467-017-00504-9

    Figure Lengend Snippet: In vivo BAFF inhibition attenuates helminth-induced lymphangiogenesis. a C57BL/6 J wild-type mice were treated with isotype control or anti-BAFF mAb (Sandy-2) and infected with Hp . The entire chain of the mLN was collected at 12 dpi and processed for flow cytometry or histological staining. b Total weight of mLN, c total cell count, d total number of B cells, e absolute number of lymphotoxin-expressing B cells, and f total FRCs (PDPN + MadCAM1 − CD31 − ) present within the mLN as determined using flow cytometry. g mLN serial cryosections showing lymphatic organization after treatment with isotype control or anti-BAFF mAbs in naive or at 12 dpi mice ( blue ; DAPI, grays ; LYVE-1 + LECs). Scale bar = 2000 μm. Images are from a single mouse and are representative of two independent experiments each including n ≥ 2 mice/group/time point. h VEGF-A and i VEGF-C in mLN tissue homogenates as determined by ELISA. j , k total LECs (PDPN + CD-31 + ) and proliferating LECs (PDPN + CD-31 + Ki-67 + ) in the mLN of naive and 12 dpi mice treated with isotype control or anti-BAFF mAbs were determined using flow cytometry. Data represent mean ± SEM and representative of two independent experiments with n = 3mice/group/time-point. Statistical analyses were performed using ANOVA, Bonferroni’s multiple comparison test and significance donated as * P

    Article Snippet: VEGF-A levels in the culture supernatant or in tissue homogenates were determined using mouse VEGF quantikine ELISA kit (R & D system, Cat No. MMV00), where as VEGF-C levels were determined using mouse VEGF-C ELISA kit (CUSABIO, CSB-E07361m) according to the manufacturer’s instructions.

    Techniques: In Vivo, Inhibition, Mouse Assay, Infection, Flow Cytometry, Cytometry, Staining, Cell Counting, Expressing, Enzyme-linked Immunosorbent Assay