Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: Increased co-expression of OSM or OSMRβ and VEGF correlates with decreased survival of IDC patients. Kaplan–Meier survival curves of invasive ductal breast carcinoma for A) OSM and VEGF (n = 318) and B) OSMRβ and VEGF expression (n = 389), as separated by quartiles. As co-expression of OSM and VEGF increases, overall survival trends decrease, with patients expressing high OSM high VEGF (upper quartiles, respectively) exhibiting significantly worse survival ( P = .0192) than individuals expressing low OSM low VEGF (lower quartiles, respectively). A comparable trend is observed with co-expression of OSMRβ and VEGF ( P = .0012). Analysis obtained from Oncomine dataset entitled Curtis Breast. Log-rank test * P < .05, ** P < .01.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

Techniques: Expressing

Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: Inflammatory cytokine and VEGF co-expression are correlated with decreased survival of invasive ductal carcinoma patient. These Kaplan–Meier curves in Figure 1, A and B , are a subset of the data presented in . A) Kaplan–Meier survival curves of invasive ductal breast carcinoma patients with high OSM and VEGF expression (upper quartiles, respectively) present diminished survival when compared to individuals with low OSM and VEGF expression (lower quartiles, respectively). Comparable trends appear upon examination of the upper and lower quartiles of patients with B) OSMRβ and VEGF expression, C) IL-6 and VEGF expression, and D) IL-6Rα and VEGF expression, indicating significant differences between upper and lower quartile survival for each group. Survival of patients with invasive ductal breast carcinoma by E) LIF and VEGF expression and F) LIFRβ and VEGF expression are not significantly different. Analysis obtained from Oncomine dataset entitled Curtis Breast. Log-rank test * P < .05, ** P < .01, *** P < .001. Inflammatory cytokine and VEGF co-expression are correlated with decreased survival of invasive ductal carcinoma patient. These Kaplan–Meier curves in Figure 1, A and B , are a subset of the data presented in Supplemental Figure S1. A) Kaplan–Meier survival curves of invasive ductal breast carcinoma patients with high OSM and VEGF expression (upper quartiles, respectively) present diminished survival when compared to individuals with low OSM and VEGF expression (lower quartiles, respectively). Comparable trends appear upon examination of the upper and lower quartiles of patients with B) OSMRβ and VEGF expression, C) IL-6 and VEGF expression, and D) IL-6Rα and VEGF expression, indicating significant differences between upper and lower quartile survival for each group. Survival of patients with invasive ductal breast carcinoma by E) LIF and VEGF expression and F) LIFRβ and VEGF expression are not significantly different. Analysis obtained from Oncomine dataset entitled Curtis Breast. Log-rank test * P < .05, ** P < .01, *** P < .001.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

Techniques: Expressing

Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: Co-expression of VEGF with OSM or OSMRβ appears to affect survival in HER2- but not HER2+ subtypes. A) Kaplan–Meier survival curves of invasive ductal breast carcinoma for HER2- patients and HER2+ patients by OSM and VEGF expression. In HER2- patients, high OSM and high VEGF expression (upper quartiles, respectively) is strongly correlated with decreased survival when compared to patients with low OSM and low VEGF expression (lower quartiles, respectively). However, this trend is not evident in HER2+ individuals, as no significant difference is observed between survival of patients with OSM and VEGF expression in upper and lower quartiles, respectively. Similar trends are observed in Kaplan–Meier survival curves for B) HER2- and HER2+ by OSMRβ and VEGF expression. Analysis obtained from Oncomine dataset entitled Curtis Breast. Log-rank test * P < .05, ** P < .01, *** P < .001.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

Techniques: Expressing

Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: IL-6 family cytokine OSM induces VEGF secretion independent of HIF1α signaling in MDA-MB-231 cells. A) Treatment with OSM, IL-6, and LIF (25 ng/mL) for 24 hours promotes expression of HIF1α in MDA-MB-231 TNBC cells. Addition of an siRNA targeting HIF1α (siHIF1α) ablates the effect of adding cytokines. B) T47D cells treated with IL-6 family cytokines for 72 hours express greater levels of HIF1α relative to non-treated control. Treatment with siHIF1α reduces this effect, as assessed by immunoblot analysis. Immunoblots are representative of at least 3 experiments. Induction of VEGF secretion is observed following treatment with IL-6 family cytokines (25 ng/mL) for 24 hours in C) MDA-MB-231 cells (n = 5) and for 72 hours in D) T47D cells (n = 3), as determined by ELISA. Treatment with siHIF1α does not affect VEGF secretion in OSM-treated MDA-MB-231 cells; however, VEGF secretion is modestly reduced in T47D cells treated with OSM and siHIF1α. E) A panel of additional human breast cancer cell lines were tested for VEGF expression by ELISA. HER2- MCF7 cells have significant OSM-induced VEGF secretion, while HER2+ BT474, SK-BR-3, and MDA-MB-453 cells do not produce increased VEGF in response to OSM. One-way ANOVA, Tukey's post-test, ** P < .01, *** P < .001.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

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

Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: OSM strongly induces phosphorylation of STAT3. A) Treatment with OSM (25 ng/mL) for 15 minutes, 30 minutes, and 1 hour strongly induces phosphorylation of STAT3 (Tyr 705) in MDA-MB-231 cells and moderately induces JNK phosphorylation (T183/Y185). Treatment with either IL-6 or LIF (25 ng/mL) does not induce pSTAT3 or pJNK. B) Treatment with OSM induces phosphorylation of STAT3 and JNK in T47D cells. IL-6 induces moderate phosphorylation of STAT3 and JNK at early time points. Phosphorylation of STAT3 and JNK was compared to control β-actin, as assessed by immunoblot analysis. Blots are representative of three experiments. Time course experiment for STAT3 phosphorylation upon treatment with OSM or IL-6 (25 ng/mL) for 0.25–72 hours in C) MDA-MB-231 cells, and in D) T47D cells. Treatment with IL-6 does not induce pSTAT3 expression in MDA-MB-231 cells, while inducing moderate pSTAT3 expression in T47D cells. ELISA was performed in quadruplicate; two-way ANOVA with Bonferroni post-test, * P < .05, ** P < .01, *** P < .001.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

Techniques: Phospho-proteomics, Control, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: OSM induces VEGF via STAT3 signaling. A) Treatment with OSM (25 ng/mL) and siSTAT3 suppresses VEGF secretion by 3-fold in MDA-MB-231 cells. B) Treatment with OSM and siSTAT3 moderately reduces VEGF secretion by T47D cells. VEGF secretion was not reduced by the addition of siRNAs targeting both JNK1 and JNK2 in either C) MDA-MB-231 or D) T47D cells. Experiments were performed in triplicate two-way ANOVA with Bonferroni post-test, * P < .05, *** P < .001.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

Techniques:

Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: Mechanistically distinct regulation of VEGF secretion in MDA-MB-231 TNBC and T47D (ER+/PR+/HER2-) cells. A) The VEGF promoter can be activated by various transcription factors. B) In MDA-MB-231 cells, OSM mediates VEGF secretion by activating the STAT3 signaling pathway downstream of the OSMR (OSMRβ + gp130). In T47D cells, OSM regulates VEGF secretion via both HIF1α and the STAT3 signaling pathway.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

Techniques:

Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: Expression of cytokines or their receptors and VEGF appears to affect survival in HER2- but not HER2+ subtypes. Kaplan–Meier survival curves of invasive ductal breast carcinoma for A) HER2- and HER2+ patients by IL-6 and VEGF expression. In HER2- patients, high IL-6 and high VEGF expression (upper quartiles, respectively) is strongly correlated with decreased survival when compared to patients with low IL-6 and low VEGF expression (lower quartiles, respectively). However, this trend is not evident in HER2+ individuals. Similar trends are observed in Kaplan–Meier survival curves between HER2- and HER2+ patients by B) IL-6Rα and VEGF expression and C) LIF and VEGF expression. D) Co-expression of LIFRβ and VEGF does not significantly affect survival in either HER2- or HER2+ patients. Analysis obtained from Oncomine dataset entitled Curtis Breast. Log-rank test * P < .05, ** P < .01, *** P < .001.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

Techniques: Expressing

Journal: Translational Oncology

Article Title: Co-Expression of VEGF and IL-6 Family Cytokines is Associated with Decreased Survival in HER2 Negative Breast Cancer Patients: Subtype-Specific IL-6 Family Cytokine-Mediated VEGF Secretion 1 2

doi: 10.1016/j.tranon.2018.10.004

Figure Lengend Snippet: Co-expression of OSM or OSMRβ and VEGF differentially affects survival based on estrogen receptor status. Kaplan–Meier survival curves of invasive ductal breast carcinoma for A) HER2−/ER- and HER2+/ER- patients and B) HER2−/ER+ and HER2+/ER+ patients by OSM and VEGF expression. In patients with either HER2+/− and ER- expression, OSM/VEGF expression levels do not significantly affect survival. However, in HER2−/ER+ patients, high OSM and high VEGF expression (upper quartiles, respectively) is strongly correlated with decreased survival when compared to patients with similar receptor status expressing low OSM and low VEGF expression (lower quartiles, respectively). Similar trends respective of receptor status are observed in Kaplan–Meier survival curves for C) HER2−/ER- and HER2+/ER- patients and D) HER2−/ER+ and HER2+/ER+ patients by OSMRβ and VEGF expression. While no correlation between OSM/VEGF expression and survival is observed in HER2+/ER+ patients, the low number of samples in these populations distorts our ability to make a robust assessment of this patient subset. Analysis obtained from Oncomine dataset entitled Curtis Breast. Log-rank test * P < .05, ** P < .01.

Article Snippet: Analysis of VEGF secretion in the conditioned media (CM) of MDA-MB-231, T47D, MCF7, SK-BR-3, MDA-MB-453, and BT474 (ATCC, Manassas, VA) breast cancer cells was assessed via VEGF ELISA according to the manufacturer's protocol (Cat# Dy293B, R&D Systems, Bethesda, MD).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Combined Inhibition of Epidermal Growth Factor Receptor and Cyclooxygenase-2 Leads to Greater Anti-tumor Activity of Docetaxel in Advanced Prostate Cancer

doi: 10.1371/journal.pone.0076169

Figure Lengend Snippet: Cells grown in 1% FBS were exposed to 0.01 μmol/L docetaxel (D), 20 μmol/L gefitinib (G) or100 μmol/LNS-398 (N), alone or in combination, or with DMSO as control for 24 h. NF-KB, MMP-9 and VEGF mRNA (A and B) and protein levels (C and D) were measured as described in Materials and Methods. Values are reported as the mean±SD of three independent experiments.

Article Snippet: Antibodies against human VEGF and MMP-9 were obtained from R&D System and Bioworld, respectively.

Techniques: Control

Journal: Neuro-Oncology

Article Title: VEGF-C sustains VEGFR2 activation under bevacizumab therapy and promotes glioblastoma maintenance

doi: 10.1093/neuonc/noy103

Figure Lengend Snippet: VEGFR2 tyrosine kinase expressed by glioblastoma cells remains active under bevacizumab therapy. (A) Western blot (WB) for VEGFR2 and tubulin in a panel of patient-derived glioblastoma cultures. (B) Quantitative RT-PCR analysis of VEGF-A and VEGFR2 expression in glioblastoma cells standardized to expression in human microvascular endothelial cells (HMVECs). Data are presented as mean ± SD, n = 3. The y -axis is log 10 transformed. (C) Viability of CPH017, IN1123, and 1966 cells treated with SU1498 (10 µM) or left unstimulated. Data are presented as mean ± SD, n = 3. Significance determined by 2-way ANOVA with Bonferroni post-test. (D) Representative WB of total and phosphorylated VEGFR2 and tubulin in CPH017 cells plated overnight in Neurobasal media without epidermal growth factor and basic fibroblast growth factor, then treated with either SU1498 (30 µM for 3 h), VEGF-A (40 ng/mL, 15 min), or the combination (30 µM SU1498 for 3 h followed by 40 ng/mL VEGF-A for 15 min). (E) Quantification of relative phosphorylated-VEGFR2/tubulin levels from WB analysis. Data are presented as mean ± SD, n = 3. Significance determined by one-way ANOVA with Dunnett’s post-test correction and ratio paired 2-sample t -test. (F) Viability of glioblastoma cells treated 7 days with bevacizumab (Bev) (0–2 mg/mL). Data are presented as mean ± SD, n = 3. Significance determined by 2-way ANOVA with Bonferroni post-test correction. (G) ELISA quantification of VEGF-A secretion by glioblastoma cells treated with Bev (0.5 mg/mL, 48 h) or IgG control. Data are presented as mean ± SD, n = 3. N.D. denotes that signal could not be detected. (H) Representative WB of total and phosphorylated VEGFR2 or tubulin in CPH017 and 1966 cells treated for 72 h with Bev (0.5 mg/mL) or IgG. * P ≤ 0.05; ** P ≤ 0.01; **** P ≤ 0.0001; NS: nonsignificant.

Article Snippet: Conditioned media were collected 48–72 hours later and subjected to Human VEGF-Quantikine enzyme-linked immunosorbent assay (ELISA) or Human VEGF-C-Quantikine ELISA (R&D Systems), following manufacturer’s instructions using a Synergy2 microplate reader (BioTek).

Techniques: Western Blot, Derivative Assay, Quantitative RT-PCR, Expressing, Transformation Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Neuro-Oncology

Article Title: VEGF-C sustains VEGFR2 activation under bevacizumab therapy and promotes glioblastoma maintenance

doi: 10.1093/neuonc/noy103

Figure Lengend Snippet: VEGF-C is expressed under bevacizumab therapy and interacts with VEGFR2 in glioblastoma cells. (A) Quantitative RT-PCR analysis of VEGF-C and VEGFR3 expression in glioblastoma cells standardized to HDLECs. Data represented as mean ± SD; n = 2. The y -axis is log 10 transformed. N.D. = not detected. (B) ELISA quantification of VEGF-C secretion by glioblastoma cells treated 72 h with bevacizumab (Bev; 0.5 mg/mL) or IgG. Data represented as mean ± SEM; n = 3. Significance determined by paired 2-sample t -test. * P ≤ 0.05. (C) Correlation of VEGF-C ELISA quantification in matched samples of blood-plasma and conditioned media (72 h) from tumor-derived cell cultures of 7 glioblastoma patients. Significance was determined by Spearman’s correlation analysis. (D) Representative confocal images showing PLA signal (red), F-actin (gray), and 4′,6′-diamidino-2-phenylindole (blue) of glioblastoma cells. Staining with only VEGF-C antibody (AB) was used as a negative control. (E) Quantification of PLA dots in glioblastoma cells. Data are represented as mean ± SEM; n > 40 cells.

Article Snippet: Conditioned media were collected 48–72 hours later and subjected to Human VEGF-Quantikine enzyme-linked immunosorbent assay (ELISA) or Human VEGF-C-Quantikine ELISA (R&D Systems), following manufacturer’s instructions using a Synergy2 microplate reader (BioTek).

Techniques: Quantitative RT-PCR, Expressing, Transformation Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Derivative Assay, Staining, Negative Control

Journal: Neuro-Oncology

Article Title: VEGF-C sustains VEGFR2 activation under bevacizumab therapy and promotes glioblastoma maintenance

doi: 10.1093/neuonc/noy103

Figure Lengend Snippet: VEGF-C regulates canonical VEGFR2 survival signaling. (A) WB analysis of downstream VEGFR2 signaling in CPH017 glioblastoma cells starved for 24 h followed by stimulation with VEGF-A (40 ng/mL) or VEGF-C (0.2 µg/mL). (B) Quantitative RT-PCR analysis of VEGF-C expression in glioblastoma cells 48 h following transfection with either siCtrl (nontargeting control) or siVEGF-C (VEGF-C targeting siRNA). Data are represented as mean ± SEM; n = 3 or 4. Significance determined with 1-sample t -tests setting the hypothetical value to 1. (C) WB for VEGF-C and tubulin in CPH017 cells 72 h following siCtrl or siVEGF-C transfection. (D) Quantitative RT-PCR analysis of VEGF-A expression in CPH017 and IN1123 cells 48 h following siCtrl or siVEGF-C transfection. Data are represented as mean ± SEM; n = 3 or 4. Significance determined with 1-sample t -tests setting the hypothetical value to 1. (E) ELISA quantification of VEGF-A in conditioned media from CPH017 glioblastoma cells transfected with siCtrl or siVEGF-C. Data are represented as mean ± SEM; n = 2. Significance determined by 1-way ANOVA with Dunnett’s post-test correction. (F) Viability of CPH017 and IN1123 glioblastoma cells transfected with siCtrl or siVEGF-C. Data are represented as mean ± SEM; n = 3–5. Significance determined by 2-way ANOVA with Bonferroni post-test correction. (G) WB of pro- and cleaved (Cl.) caspase-3 and tubulin in CPH017 and IN1123 glioblastoma cells 72 h following siCtrl or siVEGF-C transfection. (H) Viability of CPH017 and IN1123 glioblastoma cells 5 days after transfection with siCtrl or siVEGF-C alone and combined treatment with 0.5 mg/mL bevacizumab (Bev). Data are represented as mean ± SEM; n = 3. Significance determined by 1-way ANOVA with Dunnett’s post-test correction. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: Conditioned media were collected 48–72 hours later and subjected to Human VEGF-Quantikine enzyme-linked immunosorbent assay (ELISA) or Human VEGF-C-Quantikine ELISA (R&D Systems), following manufacturer’s instructions using a Synergy2 microplate reader (BioTek).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Control, Enzyme-linked Immunosorbent Assay

Journal: Archives of Virology

Article Title: Therapeutic effect of an anti-human-TNF-alpha antibody and itraconazole on feline infectious peritonitis

doi: 10.1007/s00705-020-04605-7

Figure Lengend Snippet: Changes in plasma vascular endothelial growth factor (VEGF) concentration in cats. †: The cat was euthanized because its clinical condition had reached the humane endpoint

Article Snippet: Plasma concentrations of VEGF were determined using a human VEGF ELISA Kit (R & D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques: Clinical Proteomics, Concentration Assay

Journal: Journal of Neuro-Oncology

Article Title: Identification of two distinct mesenchymal stromal cell populations in human malignant glioma

doi: 10.1007/s11060-016-2302-y

Figure Lengend Snippet: a VEGF and b PGE 2 production in vitro by two different cell populations from each of two GBMs. The CD90 − populations from GBM-47 and GBM-48 produce higher levels of both VEGF and PGE 2 compared to the CD90 + populations from the same tumors. 1 × 10 5 cells were grown in 1 ml MSC Expansion Media for 24 h and the supernatants were analyzed with ELISA. Each experiment was performed three times in duplicate and VEGF data from both tumors was pooled together

Article Snippet: VEGF levels were determined using the Human VEGF DuoSet kit (R&D Systems).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay