Journal: bioRxiv

Article Title: Vitronectin defines a differentiation-restraining extracellular matrix state in myogenic cells

doi: 10.64898/2026.01.14.699461

Figure Lengend Snippet: a, Schematic representation of the myogenic differentiation processes in C2C12 myoblasts dependent on culture medium and expression dynamics of representative muscle differentiation markers. Proliferation of myoblasts requires growth medium (GM; 10% fetal bovine serum (FBS) supplemented medium), whereas differentiation medium (DM; 2% horse serum (HS) supplemented medium) induces rapid differentiation into multinucleated myotubes and loss of proliferative capacity. b, Representative phase contrast and immunofluorescence images showing expression of the differentiation marker Myogenin on day 3 in DM. White arrowheads indicate representative differentiating myotubes. Differentiation index quantified by the proportion of myogenin-positive cells relative to total nuclei is shown in the graph. (Control: n = 1652, Vitronectin: n = 1746) c, Time-course observation of myotube differentiation by immunofluorescence for myosin heavy chain (MHC). d, Fusion index quantified by the number of nuclei per MHC-positive myotube on day 3 in DM shown in the graph (Control: n=50, Vitronectin: n=50 in MHC-positive cells). The numbers shown in the immunofluorescence images indicate of nuclei in the representative MHC-positive myotube. e, Immunoblot analysis of C2C12 cells for the representative myogenic differentiation markers: MyoD, Myogenin, and MHC. f, Immunoblot analysis of C2C12 cells for non- and phosphorylated form of STAT3, ERK1/2, and p38 MAPK. g-h , Immunofluorescence analysis of MHC expression after 3 days in DM under the indicated conditions: control, VN, VN plus cRGDfV, and cRGDfV alone. Quantification shows the proportion of MHC-positive cells relative to total nucle as differentiation indexi (g) and fusion index (h). Scale bar: 100 µm. Nuclei were counterstained with DAPI. Data in the graphs are presented as mean ± s.d. Statistical significance determined using Student’s t-test; ** p < 0.01 .

Article Snippet: Beads were then incubated with 10 μL of anti-vitronectin 65/75 antibody (D-8; Santa Cruz, sc-74484) and control antibody (Santa Cruz, sc-3877) overnight at 4°C.

Techniques: Cell Characterization, Expressing, Immunofluorescence, Marker, Control, Western Blot

Journal: bioRxiv

Article Title: Vitronectin defines a differentiation-restraining extracellular matrix state in myogenic cells

doi: 10.64898/2026.01.14.699461

Figure Lengend Snippet: a, Schematic representation of the experimental workflow to evaluate the proliferative capacity of C2C12 myoblasts cultured at low cell density in supplementation with vitronectin (GM: growth medium, DM: differentiation medium). b, Representative nuclei images stained with DAPI and the relative cell number of C2C12 cells counted on day 3 of differentiation shown in the graph (n = 6). c, Representative immunofluorescence images for phospho-Histone H3 (pH3) on day 3 of differentiation and the proportion of pH3-positive cells relative to total nuclei shown in the graph (Control: n = 1345, Vitronectin: n = 2632). d, Representative immunofluorescence images for cleaved caspase-3 on day 2 of differentiation. The ratio of apoptotic cells relative to total nuclei is shown in the graph (Control: n = 2499, Vitronectin: n = 2345). e, Western blot analysis of C2C12 cells for p21 and p27, cell cycle inhibitors. f, Immunoblot analysis of C2C12 cells for non- and phosphorylated insulin-like growth factor receptor (IGFR) and fibroblast growth factor receptor (FGFR), key growth factor receptors involved in the proliferation of C2C12 cells. Scale bar: 100 µm. White arrowheads indicate representative positive cells for p-Histone H3 (c) and cleaved caspace-3 (d). Data are presented as mean ± s.d. Statistical significance determined using Student’s t-test; ** p < 0.01 , * p < 0.05 .

Article Snippet: Beads were then incubated with 10 μL of anti-vitronectin 65/75 antibody (D-8; Santa Cruz, sc-74484) and control antibody (Santa Cruz, sc-3877) overnight at 4°C.

Techniques: Cell Culture, Staining, Immunofluorescence, Control, Western Blot

Journal: bioRxiv

Article Title: Vitronectin defines a differentiation-restraining extracellular matrix state in myogenic cells

doi: 10.64898/2026.01.14.699461

Figure Lengend Snippet: a, Schematic representation of the rotary suspension spheroid culture system. C2C12 cells were first aggregated under static conditions in a low-attachment U-shaped 96-well plate for 1 day, then transferred to suspension culture on an orbital shaker inside an incubator (37 °C, 150 rpm.) for another 5 days. b, Quantification of total cell numbers per spheroid on day 6 (n = 5). c, Representative phase-contrast and immunofluorescence images of spheroid for MHC and DAPI. Scale bar: 100 µm. d, Representative confocal images of spheroid stained for MHC, Ki-67, and DAPI on day 6. Quantification of the proportion of MHC- or Ki-67-positive cells relative to total nuclei is shown in the graphs (Control: n = 1537, Vitronectin: n = 1522). Scale bar: 50 µm. Data are presented as mean ± s.d. Statistical significance was determined using Student’s t-test; ** p < 0.01 .

Article Snippet: Beads were then incubated with 10 μL of anti-vitronectin 65/75 antibody (D-8; Santa Cruz, sc-74484) and control antibody (Santa Cruz, sc-3877) overnight at 4°C.

Techniques: Suspension, Immunofluorescence, Staining, Control

Journal: bioRxiv

Article Title: Vitronectin defines a differentiation-restraining extracellular matrix state in myogenic cells

doi: 10.64898/2026.01.14.699461

Figure Lengend Snippet: a, Immunoblot analysis of VN protein levels in fetal bovine serum, adult bovine serum, and adult horse serum. b, Immunoblot analysis of VN protein on day 0 (d0; before differentiation induction), day 1, and day 3 (d1 and d3; 1 and 3 day after differentiation induction). c, Representative immunofluorescence images showing the level and localization of VN protein in growth medium (GM) and day 1 in differentiation medium (DM). d, qPCR analysis for transcription of VN gene on d0, d1, and d3 after differentiation induction (n = 3). e, Immunoblot analysis for confirmation of immunodepleting of vitronectin from FBS. Vitronectin was depleted from FBS using control and anti-vitronectin antibody immobilized on Protein A Sepharose. f, Quantification of the inhibitory effect on myoblast differentiation with control and VN-immunodepleted FBS by immunofluorescence for MHC. (Control FBS: n = 1688, VN-reduced FBS: n = 1663 in counted nuclei number). g, Representative phase-contrast images of C2C12 cells cultured in GM, DM, and a serum-free DA-X medium supplemented with LIF on plates coated with or without VN coating. Images were captured on day 5 in each condition. Scale bar: 100 µm. Data are presented as mean ± s.d. Statistical significance was determined using one-way ANOVA in d and Student’s t-test in f ; * p < 0.05 .

Article Snippet: Beads were then incubated with 10 μL of anti-vitronectin 65/75 antibody (D-8; Santa Cruz, sc-74484) and control antibody (Santa Cruz, sc-3877) overnight at 4°C.

Techniques: Western Blot, Immunofluorescence, Control, Cell Culture

Journal: bioRxiv

Article Title: Vitronectin defines a differentiation-restraining extracellular matrix state in myogenic cells

doi: 10.64898/2026.01.14.699461

Figure Lengend Snippet: a, Representative immunofluorescence images for MHC showing myotube maturation. Maturation was quantified by the number of nuclei per MHC-positive myotube (Control: n=100, Vitronectin: n = 100 in MHC-positive cells). Scale bar: 50 µm. b, The relative cell number of chick myogenic cells cultured in indicated conditions (n = 3). c, Representative immunofluorescence images for phosphorylated histone H3 (p-Histone H3) and the proportion of positive cells relative to total nuclei (Control: n = 7619, Vitronectin: n = 7928). Scale bar: 50 µm. d, Representative phase-contrast and nuclei images of spheroid on day 5 in rotary suspension spheroid culture. Scale bar: 100 µm. In a - c , cells were analyzed 6 h after differentiation induction in DM. Nuclei were counterstained with DAPI. Data are presented as mean ± standard deviation. Statistical significance was determined using Student’s t-test; ** p < 0.01 .

Article Snippet: Beads were then incubated with 10 μL of anti-vitronectin 65/75 antibody (D-8; Santa Cruz, sc-74484) and control antibody (Santa Cruz, sc-3877) overnight at 4°C.

Techniques: Immunofluorescence, Control, Cell Culture, Suspension, Standard Deviation

Journal: PLoS ONE

Article Title: Proteomic Analysis of Mitochondrial-Associated ER Membranes (MAM) during RNA Virus Infection Reveals Dynamic Changes in Protein and Organelle Trafficking

doi: 10.1371/journal.pone.0117963

Figure Lengend Snippet: (A) Venn diagram of proteins that share a MAM-localization profile similar to that of MAVS. Proteins that move out of the MAM fraction during HCV (greater than 2-fold change over mock with a Benjamini-Hochberg corrected p-value of < 0.05) were compared to those that either move into the MAM or remain localized to the MAM during SenV infection. The 281 proteins in the intersection share this MAVS-localization pattern in the MAM during RNA virus infection. (B) Heat map of the 281 proteins with “MAVS-like” localization identified in MAM fractions from HCV and SenV, as well as mock (see panel A). Log 10 -transformed spectral counts across technical triplicates are represented by the color intensity shown in the key. (C) Immunoblot analysis of lysate (Input) and anti-Myc immunoprecipated extracts (Pellet) from cells expressing Myc-MAVS (+) or vector (-), and either Flag-RAB1B, Flag-VTN, or Flag-LONP1. (D) IFN-β promoter reporter luciferase expression of Huh7 cells expressing empty vector or Flag-LONP1 and then mock or SenV infected (24h). Values are mean -/+ SD (n = 3) of one of three replicate experiments; **P<0.01. (Inset) Immunoblot for Flag-LONP1 and tubulin (Tub.) protein expression.

Article Snippet: Plasmids encoding human VTN (NM_000638.3), LONP1 (NM_004793.2), and RAB1B (NM_030981.1) were obtained (Origene).

Techniques: Infection, Virus, Transformation Assay, Western Blot, Expressing, Plasmid Preparation, Luciferase

Journal: The Journal of biological chemistry

Article Title: The use of fluorescent probes to characterize conformational changes in the interaction between vitronectin and plasminogen activator inhibitor-1.

doi: 10.1074/jbc.272.8.5112

Figure Lengend Snippet: FIG. 1. Crystal structures that represent models for latent and active serpins. Ribbon diagrams of latent PAI-1 and ovalbumin were generated using the Insight program on a Silicon Graphics workstation. Coordinates from the x-ray crystal structures of the two proteins (20, 52) were used. b-Sheet A is visible on the surface of latent PAI-1 and has been identified as a vitronectin-binding site. The reactive center loop is shown in blue. In the active form of PAI-1, the reactive center loop is thought to be an exposed loop extending from the b-sheet on the surface of the protein, similar in structure to the homologous region shaded in blue in the ovalbumin structure. Amino acids depicted in the green and yellow space-filling models represent the P19 and P9 posi- tions, respectively. Note that the P19 and P9 residues are located within the exposed reactive center loop in the model for the active structure, and these two amino acids are buried within the hydrophobic core of the protein in the latent form of PAI-1.

Article Snippet: Murine monoclonal antibodies directed against human vitronectin were obtained from Quidel.

Techniques: Generated, Binding Assay

Journal: The Journal of biological chemistry

Article Title: The use of fluorescent probes to characterize conformational changes in the interaction between vitronectin and plasminogen activator inhibitor-1.

doi: 10.1074/jbc.272.8.5112

Figure Lengend Snippet: FIG. 2. Fluorescence changes in NBDP1*PAI-1 upon interac- tion with vitronectin. A shows the fluorescence emission spectrum (excitation wavelength 5 480 nm) for a 490 nM solution of NBDP19PAI-1 in the presence (dashed line) and absence (solid line) of native vitronectin added to a 1.3 mM final concentration. B shows binding isotherms for the interaction between NBDP19PAI-1 and native vitronectin (G), NBDP19PAI-1 and multimeric vitronectin (E), and NBDP9 PAI-1 with native vitronectin (å). Fluorescence intensity, using an excitation wavelength of 480 nm and an emission wavelength of 540 nm, was recorded following each addition of vitronectin. The data for native vitronectin interacting with NBDP19PAI-1 (G) were analyzed as described in “Experimental Procedures” to generate the binding iso- therm shown with the solid line.

Article Snippet: Murine monoclonal antibodies directed against human vitronectin were obtained from Quidel.

Techniques: Fluorescence, Concentration Assay, Binding Assay

Journal: The Journal of biological chemistry

Article Title: The use of fluorescent probes to characterize conformational changes in the interaction between vitronectin and plasminogen activator inhibitor-1.

doi: 10.1074/jbc.272.8.5112

Figure Lengend Snippet: FIG. 3. Fluorescence emission spectra of HOCGO-modified vitronectin. Native vitronectin (2 mM) was labeled with a 5:1 molar ratio of hydroxycoumarin glyoxal to protein in the presence and absence of added heparin as described under “Experimental Procedures.” The fluorescence emission spectra of the labeled products produced in the absence (solid line) and presence (dashed line) of heparin added to a 0.4 mM concentration are shown. The excitation wavelength was 335 nm.

Article Snippet: Murine monoclonal antibodies directed against human vitronectin were obtained from Quidel.

Techniques: Fluorescence, Modification, Labeling, Produced, Concentration Assay

Journal: The Journal of biological chemistry

Article Title: The use of fluorescent probes to characterize conformational changes in the interaction between vitronectin and plasminogen activator inhibitor-1.

doi: 10.1074/jbc.272.8.5112

Figure Lengend Snippet: FIG. 5. Heparin binding immunoassays comparing modified and unmodified vitronectin. Unlabeled native vitronectin (G) or HOCGO-VN (E) was serially diluted in heparin-coated microtiter plates. Bound vitronectin was detected with anti-vitronectin antibodies as described under “Experimental Procedures.” Absorbance at 405 nm (average of three measurements) is plotted versus the vitronectin con- centration (mM) expressed on a logarithmic scale.

Article Snippet: Murine monoclonal antibodies directed against human vitronectin were obtained from Quidel.

Techniques: Binding Assay, Modification

Journal: The Journal of biological chemistry

Article Title: The use of fluorescent probes to characterize conformational changes in the interaction between vitronectin and plasminogen activator inhibitor-1.

doi: 10.1074/jbc.272.8.5112

Figure Lengend Snippet: FIG. 6. PAI-1 binding activity of HOCGO-VN. HOCGO-VN (E) was assayed for the ability to compete with immobilized unmodified vitronectin for binding PAI-1. The binding activity of modified vitronec- tin is compared with the PAI-1 binding of unmodified vitronectin (G) in the same assay. The absorbance of samples in wells containing PAI-1 and no competing vitronectin provided the value for 100% binding. Percent binding inhibition was determined by subtracting the absorb- ance at each concentration of competing vitronectin from this value and then dividing by the absorbance in the absence of competing vitronectin.

Article Snippet: Murine monoclonal antibodies directed against human vitronectin were obtained from Quidel.

Techniques: Binding Assay, Activity Assay, Modification, Inhibition, Concentration Assay

Journal: The Journal of biological chemistry

Article Title: The use of fluorescent probes to characterize conformational changes in the interaction between vitronectin and plasminogen activator inhibitor-1.

doi: 10.1074/jbc.272.8.5112

Figure Lengend Snippet: FIG. 7. Kinetics of PAI-1 stabilization by HOCGO-VN. PAI-1 was incubated at 37 °C in the presence (M) or absence (G) of HOCGO-VN for periods up to 8 h to measure stabilizing effects of the vitronectin sample on PAI-1 activity. At intermediate time intervals, remaining active PAI-1 was determined by its ability to inhibit urokinase in a colorimet- ric assay, as described in “Experimental Procedures.”

Article Snippet: Murine monoclonal antibodies directed against human vitronectin were obtained from Quidel.

Techniques: Incubation, Activity Assay

Journal: The Journal of biological chemistry

Article Title: The use of fluorescent probes to characterize conformational changes in the interaction between vitronectin and plasminogen activator inhibitor-1.

doi: 10.1074/jbc.272.8.5112

Figure Lengend Snippet: FIG. 10. Assays to distinguish independent versus competitive binding of heparin and PAI-1 to vitronectin. A, serial dilutions of PAI-1 were incubated with surface-immobilized vitronectin in the ab- sence (G) or presence (E) of heparin, and bound PAI-1 was detected immunochemically as described under “Experimental Procedures.” As- says were performed using conditions in which the absorbance shows a linear relationship with the amount of PAI-1 bound to vitronectin. B, the effect of heparin on a competitive PAI-1-binding assay was deter- mined as described under “Experimental Procedures.” The percent in- hibition of PAI-1 binding observed at various concentrations of vitronec- tin is shown. The assays were performed in the presence of 0 (G), 0.4 (E), 4.0 (å), and 40 mM heparin (Ç). C, the effect of PAI-1 on the interaction between vitronectin and heparin was determined by incu- bating various concentrations of vitronectin in microtiter plates coated with 1 mgzml21 heparin. Serial dilutions of vitronectin in buffer alone (G) or in a solution containing 0.6 mM PAI-1 (E) were tested in the assay for effective binding to the heparin-coated plates.

Article Snippet: Murine monoclonal antibodies directed against human vitronectin were obtained from Quidel.

Techniques: Binding Assay, Incubation

Journal: BioTechniques

Article Title: Expression and purification of bioactive, low-endotoxin recombinant human vitronectin.

doi: 10.2144/000114181

Figure Lengend Snippet: Figure 2. Bioactivity of low-endotoxin rhVTN in HUVEC adhesion assays. (A) Promotion of HUVEC adhesion. MaxiSorp 96-well plates (Nunclon) were coated with rhVTN at the indicated concentrations (100 mL/well at 37°C for 1 h; endotoxin content 0.09 EU/mg) then blocked with heat- denatured 2% BSA. HUVECs (Life Technologies) at passage 2 or 3 were detached with Accutase, pelleted and resuspended at 3 × 105 cells/mL in warm EBM-2 (Lonza) containing 20 mM HEPES (pH 7.4 at 37°C), 1 mg/ mL hydrocortisone, 75 mM (+)-sodium-L-ascorbate, 500 mM 2-phospho- L-ascorbic acid trisodium salt, 1 mg/mL gentamicin, and 100 ng/mL heparin. Adhesion of 100 mL of HUVECs to coated wells was quantified after incubation at 37°C, 5% O2, 5% CO2, and 90% N2 for 90 min in a humidified chamber and is shown relative to that of BSA-blocked wells. Error bars depict the mean ± SEM from three independent experiments. (B) Comparison of rhVTN with other VTN sources and the effects of antagonists of integrin activity. A function-blocking mouse anti-VTN (LS- C150852, LifeSpan BioSciences, Inc.) or isotype-matched negative control MAb (50 mg/mL), EDTA (10 mM), RGES or RGDS peptide (10 mg/mL; Sigma), function-blocking mouse anti-avb3 (clone LM609, MAB1976Z; Millipore) and/or anti-avb5 (clone P1F6, MAB1961Z; Millipore) MAb, or a non-function–blocking mouse anti-avb5 MAb (clone 15F11, MAB2019Z; Millipore) were used to interrogate the mechanism of HUVEC adhesion promoted by rhVTN. Anti-integrin MAbs (15) were used at a final concentration of 20 mg/mL. The adhesion assay was performed as in panel (A) using wells coated with 1 mM rhVTN, hpVTN, or rhVTN-N (gray bars). Adhesion is expressed relative to that of rhVTN-coated wells (first bar, 100%). Bars depict the mean + SEM from three independent experiments. MAb, monoclonal antibody; FB, function-blocking; hp, human plasma.

Article Snippet: Comparison with other VTN sources at a coating concentration of 1 mM demonstrated that enhancement of HUVEC adhesion by our rhVTN was equipotent compared to commercial carrier-free hpVTN (2349- VN-100; R&D Systems, Minneapolis, MN) and similar to rhVTN-N (Figure 2B, gray bars).

Techniques: Incubation, Comparison, Activity Assay, Blocking Assay, Negative Control, Concentration Assay, Cell Adhesion Assay, Clinical Proteomics

Journal: PLoS ONE

Article Title: Functional role of vitronectin in breast cancer

doi: 10.1371/journal.pone.0242141

Figure Lengend Snippet: Displays mean fold change and standard error (SE) from the RPPA dataset, comparing race and tumor subtypes to the control group. Data allows us to compare level of vitronectin in serum. The mean fold change is relative to the control population for the same ethnicity.

Article Snippet: The vitronectin primary antibody (RD systems, MAB2349), PI3K (Cell Signaling Technology, 4292), P-AKT (Cell Signaling Technology, #9271), AKT (Cell Signaling Technology, #9272) were diluted with antibody diluent (ProteinSimple).

Techniques: Control

Journal: PLoS ONE

Article Title: Functional role of vitronectin in breast cancer

doi: 10.1371/journal.pone.0242141

Figure Lengend Snippet: A . Image displays the RPPA dot blot which was stained with the vitronectin antibody. Intensity values were taken from here and then analyzed. B-F . Displays comparisons of mean relative values of each group using standard error for error bars. B . analyzes overall serum vitronectin levels in control group (n = 40) and cancer group (n = 200). C . analyzes serum vitronectin levels (overall) in control groups and compared with the other tumor subtypes. Her2+ and LB2 show significant differences when compared to control group. D . compares control and cancer groups by ethnicity, and we find that CA cancer shows a significantly higher vitronectin concentration levels. E . we see that it compares vitronectin levels of AA and WA patients by dividing into various tumor characteristics. However, we found that there are significant differences in Her2+ and LB2 subtypes in the AA group when compared to AA controls. F . compares recurrent versus non-recurrent patient samples by ethnic groups. We found that there is a significant difference in serum vitronectin levels in non-recurrent CA compared to the control and recurrent groups, while AA samples had similar levels of serum vitronectin in all three groups.

Article Snippet: The vitronectin primary antibody (RD systems, MAB2349), PI3K (Cell Signaling Technology, 4292), P-AKT (Cell Signaling Technology, #9271), AKT (Cell Signaling Technology, #9272) were diluted with antibody diluent (ProteinSimple).

Techniques: Dot Blot, Staining, Control, Concentration Assay

Journal: PLoS ONE

Article Title: Functional role of vitronectin in breast cancer

doi: 10.1371/journal.pone.0242141

Figure Lengend Snippet: The data above is validation of data shown in , that showed potential racial disparities in tumor subtypes in vitronectin concentration levels of serum. A . Displays the relative concentration levels of vitronectin in AA Her2+ patients in serum compared with corresponding control samples. B . Relative serum concentration levels of vitronectin in WA Her2+ patients and compared with healthy patients C . Displays the relative serum concentration levels of vitronectin in AA LB2 patients and control serum. D . Displays the relative concentration levels of vitronectin in CA LB2 patients in serum.

Article Snippet: The vitronectin primary antibody (RD systems, MAB2349), PI3K (Cell Signaling Technology, 4292), P-AKT (Cell Signaling Technology, #9271), AKT (Cell Signaling Technology, #9272) were diluted with antibody diluent (ProteinSimple).

Techniques: Biomarker Discovery, Concentration Assay, Control

Journal: PLoS ONE

Article Title: Functional role of vitronectin in breast cancer

doi: 10.1371/journal.pone.0242141

Figure Lengend Snippet: A . Overall concentration level of vitronectin in 21 recurrence samples (n = 21) and compared serum vitronectin concentration levels to 21 non-recurrence samples of same IHC sub-groups. B . Receiver Operation Characteristic (ROC) Curve and, the statistical evaluation of vitronectin concentration in serum samples between non-recurrence vs recurrence patients. We tested to see if vitronectin could be used as a recurrence biomarkers and, we found that the area under curve (AUC) is 0.7107 with a p-value 0.0940.

Article Snippet: The vitronectin primary antibody (RD systems, MAB2349), PI3K (Cell Signaling Technology, 4292), P-AKT (Cell Signaling Technology, #9271), AKT (Cell Signaling Technology, #9272) were diluted with antibody diluent (ProteinSimple).

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Functional role of vitronectin in breast cancer

doi: 10.1371/journal.pone.0242141

Figure Lengend Snippet: A . Displays the pseudo blot extracted from SimpleWestern experiments. There are four cell-lines- MDA-MB-231, MCF7, MDA-MB-468, HCC1599, were used to evaluate the signaling cascade relationship. Different protein expression levels were normalized with an averaged house-keeping GAPDH and β-actin expression B . Compares vitronectin concentration levels in four BC cell lines. C . PI3K concentration levels in BC same cell-lines. D . compares AKT concentration and, E . P-AKT concentration levels in the same four BC cell lines.

Article Snippet: The vitronectin primary antibody (RD systems, MAB2349), PI3K (Cell Signaling Technology, 4292), P-AKT (Cell Signaling Technology, #9271), AKT (Cell Signaling Technology, #9272) were diluted with antibody diluent (ProteinSimple).

Techniques: Expressing, Concentration Assay

Journal: Tissue Engineering. Part A

Article Title: Vitronectin-Based, Biomimetic Encapsulating Hydrogel Scaffolds Support Adipogenesis of Adipose Stem Cells

doi: 10.1089/ten.tea.2015.0550

Figure Lengend Snippet: Effects of full-length ECM proteins on ASCs. (A) Proliferation on different substrates was quantified (*p < 0.05) (B) Secretion of adipogenic factors (adiponectin and leptin) by ASCs cultured on different ECM proteins was quantified by ELISA at day 1 (dark bars) and day 14 (light bars). Secretion of bFGF and SDF1- α (C) and VEGF and HGF (D) by ASCs cultured on different ECM proteins was quantified by ELISA over time (***p < 0.001; **p < 0.01; *p < 0.05). bFGF, basic fibroblast growth factor; Col I, collagen I; Col IV, collagen IV; FN, fibronectin; HGF, hepatocyte growth factor; LM, Laminin; SDF-1α, stromal cell-derived factor 1α; VEGF, vascular endothelial growth factor; VN, vitronectin.

Article Snippet: Laminin (L4544; Sigma-Aldrich), fibronectin (4305-FN-200; R&D systems, Minneapolis, MN), vitronectin (2308-VN-050; R&D systems), collagen type I (6220-CL; R&D Systems), and collagen type IV (CC076; Millipore) full-length proteins were used.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Derivative Assay