Journal: Veterinary Research

Article Title: Dihydromyricetin alleviates Escherichia coli lipopolysaccharide-induced hepatic injury in chickens by inhibiting the NLRP3 inflammasome

doi: 10.1186/s13567-022-01024-1

Figure Lengend Snippet: Genes and primers used in this study

Article Snippet: Anti-NLRP3 (bs-10021R, 1:500), anti-gasdermin A (bs-16331R, 1:500) and anti-caspase1-p20 (bs-10743R, 1:500) antibodies were purchased from Bioss Biotech Co. Ltd. (Beijing, China).

Techniques: Sequencing

Journal: Veterinary Research

Article Title: Dihydromyricetin alleviates Escherichia coli lipopolysaccharide-induced hepatic injury in chickens by inhibiting the NLRP3 inflammasome

doi: 10.1186/s13567-022-01024-1

Figure Lengend Snippet: DHM repressed pyroptosis in LPS-induced hepatotoxicity. A – C Changes in LDH activity, IL-18 content and IL-1β content in serum from chickens administered 0.025%, 0.05% or 0.1% DHM for 14 days followed by LPS exposure (60 mg/kg). D – H Changes in the mRNA expression levels of IL-18 (fold change in IL-18/β-actin), IL-1β (fold change in IL-1β/β-actin), gasdermin A (fold change in gasdermin A/β-actin), gasdermin E (fold change in gasdermin E/β-actin), and DFNB59 (fold change in DFNB59/β-actin) in livers from chickens administered 0.025%, 0.05% or 0.1% DHM for 14 days followed by LPS exposure (60 mg/kg). I Changes in the protein expression level of gasdermin A (fold change in gasdermin A/β-actin) in the livers of chickens administered 0.025%, 0.05% or 0.1% DHM for 14 days followed by LPS exposure (60 mg/kg). J Western blot analyses of for the expression of gasdermin A and β-actin (loading control) in total tissue lysate. Values are expressed as the mean ± SD for each group ( n = 6). ** P < 0.01 vs. the control group. # P < 0.05 and ## P < 0.01 vs. the LPS-treated group.

Article Snippet: Anti-NLRP3 (bs-10021R, 1:500), anti-gasdermin A (bs-16331R, 1:500) and anti-caspase1-p20 (bs-10743R, 1:500) antibodies were purchased from Bioss Biotech Co. Ltd. (Beijing, China).

Techniques: Activity Assay, Expressing, Western Blot

Journal: bioRxiv

Article Title: Tumor cell-organized fibronectin is required to maintain a dormant breast cancer population

doi: 10.1101/686527

Figure Lengend Snippet: a) ZR-75-1 breast cancer cell line number and proliferation (Ki67 staining) over time with serum withdrawal (red) and recovery in full serum (blue). b) Survival of breast cell lines on TCPS (TC), collagen I coated coverslips (Col), or a mixture of ECM proteins inspired by those found in the bone marrow (BM)-coated coverslips. Average time to decrease confluence from 100% to 25% (scored by visual inspection by the same observer) is displayed on the left in red, and conditions where viable cells were detected at 8 weeks in culture are labeled on the right in black. c. Best performing cell lines were cultured on TCPS for 6 weeks, and stimulated with varying media conditions (GF: growth factor cocktail, CM: conditioned medium from MSCs). Heat map shows increase in confluence over 7 days stimulation in blue. d. Ki67 quantification of BT549, and ZR-75-1 plated on TCPS and e. of HCC1954 on TCPS or collagen. Black: 10% serum control, blue: day 28 serum-free culture, green: 7 day recovery. f. Immunofluorescent staining for phospho-p38 (red; Thr180/Tyr182) and DAPI (blue) in HCC1954 on collagen coverslips after 2 and 28 days of serum-starvation (Scale bar 100 μm). g. Population quantification of HCC 1954 via co-staining of Ki67, p21, and senescence-associated β-galactosidase. h. BrdU and EdU labeling experiment schematic and results for HCC 1954 on collagen. Black: double negative, red: BrdU-positive, green: EdU-positive, blue: double BrdU- and EdU-positive.. p < 0.05 was considered statistically significant. p < 0.05 is denoted with *, ≤ 0.01 with **, ≤ 0.001 with ***, and ≤ 0.0001 with ****.

Article Snippet: ECM proteins (collagen I and natural mouse laminin were purchased from Life Technologies, human plasma fibronectin from Millipore, human collagen IV from Sigma-Aldrich, recombinant human tenascin C, osteopontin, thrombospondin-1, and vitronectin from R&D Systems, and laminin 511 and laminin 521 from Biolamina) were covalently bound to the glass surface by incubation at room temperature for three hours, then with 10 mg/cm 2 MA(PEG)24 (Thermo Fisher Scientific) for two hours to block non-specific protein adsorption, rinsed three times in PBS, and UV-sterilized prior to cell seeding.

Techniques: Staining, Labeling, Cell Culture

Journal: bioRxiv

Article Title: Tumor cell-organized fibronectin is required to maintain a dormant breast cancer population

doi: 10.1101/686527

Figure Lengend Snippet: a. Top rows, immunofluorescence for matrix proteins and DAPI in HCC 1954s on collagen in serum-free medium for 2 and 28 days. Bottom, immunofluorescence and for fibronectin in HCC 1954s grown for 28 days or serum starved for 28 days and recovered in situ for 1 week. Scale: 100 μm. b. Total MMP activity of HCC 1954 cells on collagen coverslips c. Number of nuclei resulting from reactivation of dormant cells treated with a pan MMP inhibitor for 7 days (MMPi; GM6001, 25μm) or control (serum-containing media, no inhibitor). d. HCC 1954 day 7 survival on collagen, fibronectin (FN), or HCC 1954 day 28 decellularized ECM (decell ECM) coverslips.

Article Snippet: ECM proteins (collagen I and natural mouse laminin were purchased from Life Technologies, human plasma fibronectin from Millipore, human collagen IV from Sigma-Aldrich, recombinant human tenascin C, osteopontin, thrombospondin-1, and vitronectin from R&D Systems, and laminin 511 and laminin 521 from Biolamina) were covalently bound to the glass surface by incubation at room temperature for three hours, then with 10 mg/cm 2 MA(PEG)24 (Thermo Fisher Scientific) for two hours to block non-specific protein adsorption, rinsed three times in PBS, and UV-sterilized prior to cell seeding.

Techniques: Immunofluorescence, In Situ, Activity Assay

Journal: bioRxiv

Article Title: Tumor cell-organized fibronectin is required to maintain a dormant breast cancer population

doi: 10.1101/686527

Figure Lengend Snippet: a. Experimental timeline of inhibitor dosing. Day 7 experiments were dosed continually through a day 7 endpoint; Day 28 experiments were dosed continually through a day 28 endpoint; separate cultures were established for 21 days, where inhibitor dosing was initiated for an additional 7 days (‘dose established culture’). b. HCC 1954 survival at day 7 on collagen coverslips with selected inhibitors dosed for the duration of the experiment. c. Survival of HCC 1954 cells after establishment of dormant culture for 21 days, and then subsequent dosing with inhibitors through days 21-28. d. HCC1954 survival at day 7 with inhibitors when seeded onto HCC 1954 decellularized ECM. Black: control, blue: anti- β 1 (MAB17781, 1 µg/ml), green: FAK inhibitor 14 (10 µM), purple: PD0325901 (MEK inhibitor, 10 µM), red: FR180204 (ERK inhibitor 20µM), gray: cilengitide (100 µM). p < 0.05 is denoted with *, ≤ 0.01 with **, ≤ 0.001 with ***, and ≤ 0.0001 with ****.

Article Snippet: ECM proteins (collagen I and natural mouse laminin were purchased from Life Technologies, human plasma fibronectin from Millipore, human collagen IV from Sigma-Aldrich, recombinant human tenascin C, osteopontin, thrombospondin-1, and vitronectin from R&D Systems, and laminin 511 and laminin 521 from Biolamina) were covalently bound to the glass surface by incubation at room temperature for three hours, then with 10 mg/cm 2 MA(PEG)24 (Thermo Fisher Scientific) for two hours to block non-specific protein adsorption, rinsed three times in PBS, and UV-sterilized prior to cell seeding.

Techniques:

Journal: International journal of molecular medicine

Article Title: Tanshinone IIA inhibits lipopolysaccharide‑induced inflammatory responses through the TLR4/TAK1/NF‑κB signaling pathway in vascular smooth muscle cells.

doi: 10.3892/ijmm.2019.4100

Figure Lengend Snippet: Figure 2. Tan IIA inhibits LPS‑induced VSMC phenotypic switching. VSMCs were pretreated with the indicated concentrations of Tan IIA (25, 50 and 100 µmol/l) for 1 h, and then stimulated with LPS (1 µg/ml) for a further 24 (for western blotting) or 6 h (for RT‑qPCR). The (A) protein and (B) mRNA expression of α‑SMA and OPN were measured by western blotting and RT‑qPCR, respectively. Data are presented as the mean ± standard error of the mean of six independent experiments. #P<0.001 vs. control; **P<0.01 and ***P<0.001 vs. LPS. Tan IIA, Tanshinone IIA; VSMCs, vascular smooth muscle cells; LPS, lipopolysaccharide; α‑SMA, α‑smooth muscle actin; OPN, osteopontin; RT‑qPCR, reverse transcription‑quantitative polymerase chain reaction.

Article Snippet: Antibodies against TLR4, inducible nitric oxide (NO) synthase (iNOS), TAK1, phosphorylated (p)‐TAK1, α-SMA, osteopontin (OPN), NF-κB (p65), p47phox, IκBα, p-IκBα, anti-rabbit horseradish peroxidase (HRP)‐conjugated, anti‐mouse HRP‐conjugated and histone were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Western Blot, Expressing, Control, Polymerase Chain Reaction

Journal: Science Advances

Article Title: Tumor cell–organized fibronectin maintenance of a dormant breast cancer population

doi: 10.1126/sciadv.aaz4157

Figure Lengend Snippet: ( A ) ZR-75-1 breast cancer cell line number and proliferation (Ki67 staining) over time with serum withdrawal (red) and recovery in full serum (blue). ( B ) Survival of breast cell lines on TCPS (TC), collagen I coverslips (Col), or a mixture of ECM proteins inspired by those found in the bone marrow (BM)–functionalized coverslips. Average time to decrease confluence from 100 to 25% (scored by visual inspection by the same observer) is displayed on the left in red, and conditions where viable cells were detected at 8 weeks in culture are labeled on the right in black. ( C ) Best-performing cell lines were cultured on TCPS for 6 weeks and stimulated with varying media conditions (GF, growth factor cocktail; CM, conditioned medium from mesenchymal stem cells). SF, serum-free. Heat map shows increase in confluence over 7-day stimulation in blue. ( D ) Ki67 quantification of BT549 and ZR-75-1 plated on TCPS and ( E ) of HCC 1954 on TCPS or collagen. Black, 10% serum control; blue, day 28 serum-free culture; green, 7-day recovery. ( F ) Immunofluorescent staining for phospho-p38 (red; Thr 180 /Tyr 182 ) and 4′,6-diamidino-2-phenylindole (DAPI) (blue) in HCC 1954 on collagen coverslips after 2 and 28 days of serum starvation (scale bar, 100 μm). ( G ) Population quantification of HCC 1954 via costaining of Ki67, p21, and senescence-associated β-galactosidase (β-gal). ( H ) BrdU- and EdU-labeling experiment schematic and results for HCC 1954 on collagen. Black, double negative; red, BrdU positive; green, EdU positive; blue, double BrdU and EdU positive. P < 0.05 was considered statistically significant. P < 0.05 is denoted with * P < 0.05, and **** P ≤ 0.0001.

Article Snippet: ECM proteins (collagen I and natural mouse laminin were purchased from Life Technologies; human plasma fibronectin from Millipore; human collagen IV from Sigma-Aldrich; recombinant human tenascin C, osteopontin, thrombospondin-1, and vitronectin from R&D Systems; and laminin 511 and laminin 521 from BioLamina) were covalently bound to the glass surface by incubation at room temperature for 3 hours and then with methacrylate-PEG-amine [MA(PEG)] (10 mg/cm 2 ) (Thermo Fisher Scientific) for 2 hours to block nonspecific protein adsorption, rinsed three times in phosphate-buffered saline (PBS), and UV-sterilized before cell seeding.

Techniques: Staining, Labeling, Cell Culture, Control

Journal: Science Advances

Article Title: Tumor cell–organized fibronectin maintenance of a dormant breast cancer population

doi: 10.1126/sciadv.aaz4157

Figure Lengend Snippet: ( A ) Top, immunofluorescence for matrix proteins and DAPI in HCC 1954 on collagen in serum-free medium for 2 and 28 days. Bottom, immunofluorescence and for fibronectin in HCC 1954s grown for 28 days or serum-starved for 28 days and recovered in situ for 1 week (blue, DAPI; green, fibronectin; pink, laminin; red, collagen I, vitronectin, or osteopontin, as labeled). Scale bar, 100 μm. ( B ) Total MMP activity of HCC 1954 cells on collagen coverslips ( C ) Number of nuclei resulting from reactivation of dormant cells treated with a pan MMP inhibitor for 7 days (MMPi; GM6001, 25 μM) or control (serum-containing media, no inhibitor). ( D ) HCC 1954 day 7 survival on collagen, fibronectin (FN), or HCC 1954 day 28 decellularized ECM (decell ECM) coverslips.

Article Snippet: ECM proteins (collagen I and natural mouse laminin were purchased from Life Technologies; human plasma fibronectin from Millipore; human collagen IV from Sigma-Aldrich; recombinant human tenascin C, osteopontin, thrombospondin-1, and vitronectin from R&D Systems; and laminin 511 and laminin 521 from BioLamina) were covalently bound to the glass surface by incubation at room temperature for 3 hours and then with methacrylate-PEG-amine [MA(PEG)] (10 mg/cm 2 ) (Thermo Fisher Scientific) for 2 hours to block nonspecific protein adsorption, rinsed three times in phosphate-buffered saline (PBS), and UV-sterilized before cell seeding.

Techniques: Immunofluorescence, In Situ, Labeling, Activity Assay, Control

Journal: Science Advances

Article Title: Tumor cell–organized fibronectin maintenance of a dormant breast cancer population

doi: 10.1126/sciadv.aaz4157

Figure Lengend Snippet: ( A ) Experimental timeline of inhibitor dosing. Day 7 experiments were dosed continually through a day 7 endpoint; day 28 experiments were dosed continually through a day 28 endpoint; separate cultures were established for 21 days, where inhibitor dosing was initiated for an additional 7 days (dose-established culture). ( B ) HCC 1954 survival at day 7 on collagen coverslips with selected inhibitors dosed for the duration of the experiment. ( C ) Survival of HCC 1954 cells after establishment of dormant culture for 21 days and then subsequent dosing with inhibitors through days 21 to 28. ( D ) HCC 1954 survival at day 7 with inhibitors when seeded onto HCC 1954 decellularized ECM. Black, control; blue, anti-β 1 (MAB17781; 1 μg/ml); green, FAK inhibitor 14 (10 μM); purple, PD0325901 (MEK inhibitor, 10 μM); red, FR180204 (ERK inhibitor, 20 μM); gray, cilengitide (100 μM). P < 0.05 is denoted with *, P ≤ 0.001 with *** and P ≤ 0.0001 with ****.

Article Snippet: ECM proteins (collagen I and natural mouse laminin were purchased from Life Technologies; human plasma fibronectin from Millipore; human collagen IV from Sigma-Aldrich; recombinant human tenascin C, osteopontin, thrombospondin-1, and vitronectin from R&D Systems; and laminin 511 and laminin 521 from BioLamina) were covalently bound to the glass surface by incubation at room temperature for 3 hours and then with methacrylate-PEG-amine [MA(PEG)] (10 mg/cm 2 ) (Thermo Fisher Scientific) for 2 hours to block nonspecific protein adsorption, rinsed three times in phosphate-buffered saline (PBS), and UV-sterilized before cell seeding.

Techniques: Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Osteopontin and iCD8α cells promote intestinal intraepithelial lymphocyte homeostasis

doi: 10.4049/jimmunol.1901168

Figure Lengend Snippet: Osteopontin-deficient mice have a reduced IEL compartment. (A) Gating strategy utilized in this report. After gating on live cells, IEL were gated as indicated. (B) Total IEL numbers from the small intestine (top) and colon (bottom) of WT and Spp-1−/− mice. Each dot represents an individual mouse (n = 13). (C) Total cell numbers of the indicated populations in the spleens from WT and Spp-1−/− mice (n = 8). Bars indicate SEM. (D) Total cell numbers of the indicated populations in the lamina propria from WT and Spp-1−/− mice. Each dot represents an individual mouse (n = 5). Data from (B) to (D) are representative of two to three independent experiments. * P<0.05; **P<0.01; ****P<0.0001 (Mann-Whitney U test).

Article Snippet: Osteopontin and GADPH primers were from Qiagen.

Techniques: MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Osteopontin and iCD8α cells promote intestinal intraepithelial lymphocyte homeostasis

doi: 10.4049/jimmunol.1901168

Figure Lengend Snippet: Differential apoptosis and cell division in IEL from osteopontin-deficient mice. (A) Annexin V staining of different small intestine IEL populations derived from WT and Spp-1−/− mice. After gating cells by size based on forward and side scatter profiles, cells were gated for the expression of CD45 and other surface markers as indicated in Fig. 1A, and then analyzed for annexin V and 7AAD staining. Dot plots show a representative sample. Summary is indicated in the graphs. Data are representative of three independent experiments. Each dot represents an individual sample (n = 8 to 10). (B) Ki67 intracellular staining of different small intestine IEL populations derived from WT and Spp-1−/− mice. Cells were gated as in Figure 1A. Data are representative from two independent experiments. Each dot represents an individual sample (n = 9 to 10). *P<0.05; **P<0.01 (Mann-Whitney U test).

Article Snippet: Osteopontin and GADPH primers were from Qiagen.

Techniques: Staining, Derivative Assay, Expressing, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Osteopontin and iCD8α cells promote intestinal intraepithelial lymphocyte homeostasis

doi: 10.4049/jimmunol.1901168

Figure Lengend Snippet: Osteopontin promotes in vitro IEL survival. (A) Indicated FACS-enriched IEL populations from WT mice were left untreated or treated with recombinant murine osteopontin (rOPN; 2 μg/ml), with or without anti-mouse CD44 (5 μg/ml). After the indicated time points, cell survival was determined. Data are representative of three independent experiments. Biological replicas consisted of two to three pooled IEL preparations from individual mice; each experiment consisted of 3 biological replicas. (B) Enriched CD45+ splenocytes were treated as in (A). Data are representative of two independent experiments (n = 3). (C) FACS-enriched TCRβ+CD44+ spleen cells from WT and Spp-1−/− mice were treated as in (A). Data are representative of two independent experiments (n = 3). (D) Neonatal human IEL were incubated in the presence or absence of recombinant human osteopontin (2 μg/ml) and anti-human CD44 (5μg/ml). After the indicated time points, cell survival was determined. Each symbol represents an individual human: circle, small intestine from 1-day old patient presenting volvulus and necrosis; square, ileum and colon from 17-day old patient presenting necrotizing enterocolitis; triangle, jejunum from 12-day old patient presenting necrotizing enterocolitis; diamond and hexagon, de-identified individuals. (E) PBMC from adult humans were treated as in (D). Data are representative of two independent experiments (n = 3). *P<0.05, **P<0.01, ***P<0.001 (One-way ANOVA).

Article Snippet: Osteopontin and GADPH primers were from Qiagen.

Techniques: In Vitro, Recombinant, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Osteopontin and iCD8α cells promote intestinal intraepithelial lymphocyte homeostasis

doi: 10.4049/jimmunol.1901168

Figure Lengend Snippet: Environmental osteopontin influences IEL reconstitution and colon inflammation. Enriched total spleen T cells from WT mice were adoptively transferred into Rag-2−/− and Spp-1−/−Rag-2−/− mice. (A) Gating strategy to identify cells derived from donor mice (top). Representative analysis of mice analyzed seven days after transfer (bottom). Dot plots show a representative sample. Results are summarized in the graphs. TCRβ+CD4+CD8α+ cells were not detected above background at this time point in either group. Data are representative from two independent experiments. Each dot represents an individual sample (n = 4 to 5). (B) The same analysis as in (A) performed at 28 days post transfer. Dot plots show a representative sample. Results are summarized in the graphs. Data are representative from three independent experiments. Each dot represents an individual animal (n = 9). (C) Total weight change at 28 days post transfer of mice treated as in (B). (D) Representative H&E stained samples of colon sections from the indicated mice at 28 days (100X magnification). Graph indicates total histological score [immune cell infiltration (3 points), loss of goblet cells (3 points), crypt damage (3 points) and epithelial hyperplasia (3 points)]. For (C) and (D) data are representative from three independent experiments, each dot represents an individual mouse (n = 9). (E) Total mRNA expression from colons of naïve and T cell treated (28 days post-transfer) Rag-2−/− and Spp-1−/−Rag-2−/− mice (n=3–5). **P<0.01 (Mann-Whitney T test).

Article Snippet: Osteopontin and GADPH primers were from Qiagen.

Techniques: Derivative Assay, Staining, Expressing, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Osteopontin and iCD8α cells promote intestinal intraepithelial lymphocyte homeostasis

doi: 10.4049/jimmunol.1901168

Figure Lengend Snippet: Osteopontin sustains Foxp3 expression. (A) Intestinal T cells from RFP-Foxp3 reporter mice were isolated and cultured in the presence or absence of recombinant osteopontin (2 μg/ml) and anti-CD44 (5 μg/ml). Seventy-two hours later, the percentage of RFP+ cells was determined by flow cytometry. After excluding dead cells, dot plots were gated as CD45+TCRβ+ cells. (B) Bar graph indicates the fold change in CD4+RFP+ cells in relation to the null group. Data are representative of three independent experiments. (C) Enriched splenic RFP+ cells from RFP-Foxp3 reporter mice were adoptively transferred into Rag-2−/− and Spp-1−/−Rag-2−/− recipient mice. Eight weeks after transfer, IEL from small intestine were isolated and the percentage of RFP+ determined. After excluding dead cells, plots were gated as indicated by the arrows. (D) Summary of the percentage of donor-derived cells recovered; each dot represents an individual mouse. Data are representative from three independent experiments. (E) Summary of the percentage of RFP+ cells within the donor-derived cells. *P<0.05 (Mann-Whitney T test). rOPN = recombinant osteopontin; aCD44 = anti-CD44 antibodies.

Article Snippet: Osteopontin and GADPH primers were from Qiagen.

Techniques: Expressing, Isolation, Cell Culture, Recombinant, Flow Cytometry, Derivative Assay, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Osteopontin and iCD8α cells promote intestinal intraepithelial lymphocyte homeostasis

doi: 10.4049/jimmunol.1901168

Figure Lengend Snippet: iCD8α cell-derived osteopontin promotes IEL survival. Enriched CD45+ IEL from small intestine and colon derived from Spp-1−/− mice were incubated in the presence or absence of enriched iCD8α IEL (A), TCRβ+ IEL (B), or TCRγδ+ IEL (C) from osteopontin-competent donors. Some cells were cultured in the presence of anti-osteopontin antibodies (2 μg/ml). Four hours after incubation, cells were stained for surface markers, including annexin V, and 7AAD. Cells were gated as in Figure 2A. Graphs indicate “increased survival”, which was determined as 100 - [% of annexin V+ read-out cells in co-culture x 100 / % of annexin V+ cells cultured alone].

Article Snippet: Osteopontin and GADPH primers were from Qiagen.

Techniques: Derivative Assay, Incubation, Cell Culture, Staining, Co-Culture Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Osteopontin and iCD8α cells promote intestinal intraepithelial lymphocyte homeostasis

doi: 10.4049/jimmunol.1901168

Figure Lengend Snippet: IEC-derived osteopontin does not promote IEL homeostasis. Small intestine (A) and colon (B) IEL were isolated from Villin-Cre−/−Spp-1fl/fl and Villin-Cre+/−Spp-1fl/fl mice, and the cellularity determined as indicated in Fig. 1A and ​andBB.

Article Snippet: Osteopontin and GADPH primers were from Qiagen.

Techniques: Derivative Assay, Isolation

Journal: Journal of Clinical Oncology

Article Title: Development of a Multimarker Assay for Early Detection of Ovarian Cancer

doi: 10.1200/JCO.2008.19.2484

Figure Lengend Snippet: Multiplexed Biomarkers

Article Snippet: Millipore/Linco , Insulin, 64 osteopontin, 56 osteoprotegerin, 65 osteocalcin 66 , 17.

Techniques: Luminex