Journal: Theranostics

Article Title: Integrin α v β 3 -Targeted Radiotracer 99m Tc-3P-RGD 2 Useful for Noninvasive Monitoring of Breast Tumor Response to Antiangiogenic Linifanib Therapy but not Anti-Integrin α v β 3 RGD 2 Therapy

doi: 10.7150/thno.6989

Figure Lengend Snippet: A : The 3D (upper panel) and transverse (lower panel) views of SPECT/CT images of a mouse with bearing MDA-MB-435 breast cancer xenografts at 5, 7, 14, 21, 28 and 35 days after inoculation of MDA-MB-435 human breast cancer cells. The animal was administered with 37 - 55.5 MBq of 99m Tc-3P-RGD 2 via the lateral tail vein. B : Microscopic images (Original magnification: ×200) of tumor slice selected from the necrotic and viable regions after immunohistochemical staining for integrin β 3 and CD31. The tumor tissue was harvested on day 28 days after inoculation of MDA-MB-435 cells. CD31 was used as a biomarker for endothelial cells on blood vessels (both mature vasculature and neovasculture). Integrin β 3 was visualized with Cy3 (red) and CD31 was visualized with FITC (green). The yellow color in overlay images indicates the presence of integrin β 3 on neovasculature.

Article Snippet: Sections were blocked with 10% goat serum for 30 min at room temperature, and then were incubated with the hamster anti-integrin β 3 antibody (1:100, BD Biosciences, San Jose, CA) and rat anti-CD31 antibody (1:100, V:V; BD Biosciences) for 1 h. After incubating with Cy3-conjugated goat anti-hamster and FITC-conjugated goat anti-rat secondary antibodies (1:100, Jackson ImmunoResearch Inc., West Grove, PA) and washing with PBS, the fluorescence was visualized with a Nikon fluorescence microscope.

Techniques: Single Photon Emission Computed Tomography, Immunohistochemical staining, Staining, Biomarker Assay

Journal: Theranostics

Article Title: Integrin α v β 3 -Targeted Radiotracer 99m Tc-3P-RGD 2 Useful for Noninvasive Monitoring of Breast Tumor Response to Antiangiogenic Linifanib Therapy but not Anti-Integrin α v β 3 RGD 2 Therapy

doi: 10.7150/thno.6989

Figure Lengend Snippet: Microscopic images of frozen tumor slices from the xenografted MDA-MB-435 tumors (0.05 g, 0.13 g, 0.28 g, 0.55 g and 0.88 g) after immunohistochemical staining for integrin β 3 and CD31. CD31 was used as a biomarker for tumor endothelial cells. Integrin β 3 was visualized with Cy3 (red), and CD31 with FITC (green). Yellow color indicates the integrin β 3 on new blood vessels.

Article Snippet: Sections were blocked with 10% goat serum for 30 min at room temperature, and then were incubated with the hamster anti-integrin β 3 antibody (1:100, BD Biosciences, San Jose, CA) and rat anti-CD31 antibody (1:100, V:V; BD Biosciences) for 1 h. After incubating with Cy3-conjugated goat anti-hamster and FITC-conjugated goat anti-rat secondary antibodies (1:100, Jackson ImmunoResearch Inc., West Grove, PA) and washing with PBS, the fluorescence was visualized with a Nikon fluorescence microscope.

Techniques: Immunohistochemical staining, Staining, Biomarker Assay

Journal: Theranostics

Article Title: Integrin α v β 3 -Targeted Radiotracer 99m Tc-3P-RGD 2 Useful for Noninvasive Monitoring of Breast Tumor Response to Antiangiogenic Linifanib Therapy but not Anti-Integrin α v β 3 RGD 2 Therapy

doi: 10.7150/thno.6989

Figure Lengend Snippet: A : Comparison of tumor volumes in vehicle and linifanib-treated in the MDA-MB-435 models. Tumor volume was determined by caliper measurements. *: p < 0.05, significantly different from the vehicle-treated group. #: p < 0.01, significantly different from the vehicle-treated group from that specific date until the end of study. B : Transverse views of selected SPECT/CT images from athymic nude mice bearing MDA-MB-435 breast tumors in the vehicle (upper panel) and linifanib-treated groups (lower panel). SPECT/CT images were obtained at -1, 1, 4 and 11 days after initiation of linifanib therapy to illustrate the early tumor response in terms of tumor uptake and integrin a v β 3 expression levels. Arrows indicate the presence of tumors.

Article Snippet: Sections were blocked with 10% goat serum for 30 min at room temperature, and then were incubated with the hamster anti-integrin β 3 antibody (1:100, BD Biosciences, San Jose, CA) and rat anti-CD31 antibody (1:100, V:V; BD Biosciences) for 1 h. After incubating with Cy3-conjugated goat anti-hamster and FITC-conjugated goat anti-rat secondary antibodies (1:100, Jackson ImmunoResearch Inc., West Grove, PA) and washing with PBS, the fluorescence was visualized with a Nikon fluorescence microscope.

Techniques: Single Photon Emission Computed Tomography, Expressing

Journal: Theranostics

Article Title: Integrin α v β 3 -Targeted Radiotracer 99m Tc-3P-RGD 2 Useful for Noninvasive Monitoring of Breast Tumor Response to Antiangiogenic Linifanib Therapy but not Anti-Integrin α v β 3 RGD 2 Therapy

doi: 10.7150/thno.6989

Figure Lengend Snippet: Comparison of changes in %ID/cm 3 tumor uptake ( A ) and T/M ratios ( B ) of 99m Tc-3P-RGD 2 after linifanib treatment in three different tumor-bearing animal models (U87MG: high integrin α v β 3 expression on tumor cells and neovasculature; MDA-MB-435: moderate integrin α v β 3 expression on tumor cells and neovasculature; PC-3: low integrin α v β 3 expression on tumor cells and neovasculature). Tumor uptake values were calculated from quantification of SPECT/CT images. The %ID/cm 3 tumor uptake and T/M ratios of 99m Tc-3P-RGD 2 in the U87MG and PC-3 models were obtained from our previous report . The changes in %ID/cm 3 tumor uptake and T/M ratios were calculated by subtracting the value on a specific date (+1, +4 and +11) from the value on day -1. *: p < 0.05, significantly different from those in the U87MG and PC-3 models. # : p < 0.01, significantly different from those in the U87MG and MDA-MB-435 models.

Article Snippet: Sections were blocked with 10% goat serum for 30 min at room temperature, and then were incubated with the hamster anti-integrin β 3 antibody (1:100, BD Biosciences, San Jose, CA) and rat anti-CD31 antibody (1:100, V:V; BD Biosciences) for 1 h. After incubating with Cy3-conjugated goat anti-hamster and FITC-conjugated goat anti-rat secondary antibodies (1:100, Jackson ImmunoResearch Inc., West Grove, PA) and washing with PBS, the fluorescence was visualized with a Nikon fluorescence microscope.

Techniques: Expressing, Single Photon Emission Computed Tomography

Journal: Theranostics

Article Title: Integrin α v β 3 -Targeted Radiotracer 99m Tc-3P-RGD 2 Useful for Noninvasive Monitoring of Breast Tumor Response to Antiangiogenic Linifanib Therapy but not Anti-Integrin α v β 3 RGD 2 Therapy

doi: 10.7150/thno.6989

Figure Lengend Snippet: A : Overlay images of MDA-MB-435 tumor tisues after immunohistochemical staining for integrin β 3 and CD31 in vehicle (upper panel) and linifanib-treated groups (lower panel) to illustrate the changes in integrin β 3 expression level and blood vessel density during linifanib therapy. Tumor tissues were harvested at -1, 1, 4 and 11 days after initiation of linifanib therapy. CD31 was the marker for tumor blood vessels (both mature and newly formed), which was visualized with FITC (green color). Integrin β 3 was visualized with Cy3 (red color). The yellow color in overlay images indicates the presence of integrin β 3 on new blood vessels. B : Images of selected histological slice (H&E stained) of tumor tissues from animals in the vehicle (upper panels) and linifanib-treated (lower panels) groups to illustrate vascular density changes after only one day of linifanib therapy. Red color indicates the presence of blood vessels.

Article Snippet: Sections were blocked with 10% goat serum for 30 min at room temperature, and then were incubated with the hamster anti-integrin β 3 antibody (1:100, BD Biosciences, San Jose, CA) and rat anti-CD31 antibody (1:100, V:V; BD Biosciences) for 1 h. After incubating with Cy3-conjugated goat anti-hamster and FITC-conjugated goat anti-rat secondary antibodies (1:100, Jackson ImmunoResearch Inc., West Grove, PA) and washing with PBS, the fluorescence was visualized with a Nikon fluorescence microscope.

Techniques: Immunohistochemical staining, Staining, Expressing, Marker

Journal: Theranostics

Article Title: Integrin α v β 3 -Targeted Radiotracer 99m Tc-3P-RGD 2 Useful for Noninvasive Monitoring of Breast Tumor Response to Antiangiogenic Linifanib Therapy but not Anti-Integrin α v β 3 RGD 2 Therapy

doi: 10.7150/thno.6989

Figure Lengend Snippet: Linear relationship between the %ID/cm 3 tumor uptake change at days 1 (top), 4 (middle) and 11 (bottom) after linifanib therapy and the expression levels of integrin α v β 3 ( left ) and CD31 ( right ) in three animal models. The %ID/cm 3 tumor uptake values of 99m Tc-3P-RGD 2 were calculated from SPECT/CT quantification, and reported as the mean plus/minus standard error of the mean based on results from five animals (n = 5). The %ID/cm 3 tumor uptake values and integrin α v β 3 /CD31 expression levels in the U87MG and PC-3 tumors were obtained from our previous report . The %ID/cm 3 tumor uptake change was calculated by deducting the %ID/cm 3 tumor uptake of 99m Tc-3P-RGD 2 on days 1, 4 and 11 from its original value on -1 day (before linifanib therapy) in the same animal. The average %ID/cm 3 tumor uptake change is used as an indicator of tumor response to linifanib antiangiogenesis treatment.

Article Snippet: Sections were blocked with 10% goat serum for 30 min at room temperature, and then were incubated with the hamster anti-integrin β 3 antibody (1:100, BD Biosciences, San Jose, CA) and rat anti-CD31 antibody (1:100, V:V; BD Biosciences) for 1 h. After incubating with Cy3-conjugated goat anti-hamster and FITC-conjugated goat anti-rat secondary antibodies (1:100, Jackson ImmunoResearch Inc., West Grove, PA) and washing with PBS, the fluorescence was visualized with a Nikon fluorescence microscope.

Techniques: Expressing, Single Photon Emission Computed Tomography

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: Integrin expression profiles are altered after OC-2 cell differentiation . (A) Western blot analysis of myosin VI and myosin VIIa levels in OC-2 cells grown at 33°C and 39°C. Both myosin VI and VIIa levels were increased significantly in cells grown at 39°C cells in comparison with those incubated at 33°C. Bar graphs represent densitometric results of mean relative values of myosin VI and myosin VIIa levels ± s.e.m. HSC-70 provided the loading control. (B) Undifferentiated (33°C) and differentiated (39°C) OC-2 cells were analysed by FACS for the integrin subunits α6, β1, αv and β3. Levels of β1-, αv- and β3-integrin subunits were increased significantly in differentiated OC-2 cells when compared with undifferentiated OC-2 cells. α6-integrin surface expressions levels did not change between the two cell phenotypes. Bar graph represents mean fluorescence units of the various integrin subunits ± s.e.m.; n = 3 independent experiments. White bars = cells at 33°C, black bars = cells at 39°C cells; nd = no significant difference, * P < 0.05, ** P < 0.01.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Cell Differentiation, Western Blot, Incubation, Fluorescence

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: Integrin expression profiles in the Organ of Corti . RT-PCR of RNA isolated from organ of Corti of adult mice. Bands at the predicted molecular weight are visible for integrin β1, β3, αv, α6 and β-actin but not for integrin β4. cDNA was omitted for the negative control.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Molecular Weight, Negative Control

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: αv-integrin expression profiles change during differentiation . Expression of (A) α6-integrin and (B) αv-integrin were examined by FACS at days 2, 6, 10 and 14 during the differentiation process. The differentiating cells ('dif' red line) were compared to undifferentiated ('33' green line) and fully differentiated cells ('39' red line). α6-integrin expression profiles did not change throughout the time course. αv-integrin expression profiles show an increase at day 6 and are at comparable levels with fully differentiated OC-2 cells at day 14 with an apparent even higher level at day 10.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: β1 and β3-integrin expression profiles change during differentiation . Surface expression of (A) β1-integrin and (B) β3-integrin was examined by FACS at days 2, 6, 10 and 14 of the differentiation process The differentiating cells ('dif' red line) were compared to undifferentiated ('33' green line) and fully differentiated cells ('39' red line). Increase of β1-integrin expression profile is visible from day 2 and it reaches the level of differentiated cells by day 10. β3-integrin expression which is undetectable above the Ig signal up to day 2, becomes visible at day 6 and gradually reaches the differentiated levels at day 14.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: β3-integrin is a marker for OC-2 cell differentiation . (A) Western blot analysis of myosin VIIa expression in 39°C OC-2 cells untreated or after treatment with either scrambled (Scr), β1-, α6-, αv- and β3-integrin siRNA. Treatment with all siRNAs reduced myosin VIIa levels significantly when compared with untreated OC-2 cells or OC-2 cells treated with scrambled siRNA. (B) Undifferentiated OC-2 cells transduced with human β3-integrin (+hβ3) had similar levels of surface β3-integrin to differentiated (39) OC-2 cells. Furthermore, analysis of the expression of myosin VIIa showed it was increased significantly in OC-2 cells transduced with human β3-integrin (+hβ3) cells compared with undifferentiated (33) cells. Bar graphs represent densitometric results of mean relative values of myosin VIIa levels ± s.e.m. HSC-70 was used as a loading control. * P < 0.05, ** P < 0.01. n = 3 individual experiments.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Marker, Cell Differentiation, Western Blot, Expressing, Transduction

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: Effect of β3-integrin-siRNA treatment on integrin surface expression . Differentiated OC2 cells were treated with β3-siRNA and assayed by FACS for expression of surface integrins. The data is compared to integrin expression levels in normal differentiated cells (39°), differentiated cells exposed to scrambled siRNA sequences (siRNAβ3) and undifferentiated cells (33°). Exposure to β3-integrin-siRNA resulted in a significant reduction of the surface expression of β3, β1- and αv- to levels similar to those of undifferentiated cells. There was no significant effect of scrambled siRNA sequences on any integrin expression and β3-integrin-siRNA did not significantly affect α6-integrin providing further evidence for the specificity of the siRNA inhibition.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Inhibition

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: Graph of changes in expression levels of integrin subunits and myosin VIIa during OC-2 cell differentiation . Schematic time course graph showing fluorescence values of αv-, α6-, β1- and β3- integrin subunit surface expression measured by FACS and chemiluminescence values of the hair cell marker myosin VIIa in OC-2 cells measured by Western blot, over the 14 day differentiation process.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Cell Differentiation, Fluorescence, Marker, Western Blot

Journal: The Journal of Cell Biology

Article Title: Inhibition of β 2 Integrin–Mediated Leukocyte Cell Adhesion by Leucine–Leucine–Glycine Motif–Containing Peptides

doi:

Figure Lengend Snippet: Seven Phage Sequences Bound to the α M β 2 Integrin (Mac-1) and their Alignment with LLG-containing Sequences Present in Cell Adhesion Proteins

Article Snippet: The α IIb β 3 integrin antibody P2 was purchased from Immunotech, and the α V β 3 integrin antibody LM609 and the β 1 subunit antibody 6S6 were from Chemicon.

Techniques:

Journal: The Journal of Cell Biology

Article Title: Inhibition of β 2 Integrin–Mediated Leukocyte Cell Adhesion by Leucine–Leucine–Glycine Motif–Containing Peptides

doi:

Figure Lengend Snippet: Divalent cation–dependent binding of LLG-C4 nonapeptide to leukocyte β 2 integrin and its I domain. (A) Integrin from a blood cell lysate was immunocaptured on microtiter wells using the α M subunit antibody MEM170 or OKM1, the α X subunit antibody TS2/4, or the β 2 subunit antibody 2E7. Purified LLG-C4-GST or GST control (2 μg/well) was allowed to bind for 60 min in the absence or presence of EDTA. The bound GST protein was determined by using anti-GST antibodies. The results show the means ± SD from triplicate wells. The experiment was repeated three times with similar results. (B) LLG-C4-GST or GST was incubated in microtiter wells coated with purified α M subunit I domain. The concentrations of GST proteins were as indicated. The bound GST was determined with anti-GST antibodies. The results are means ± SD from triplicate wells. The results were similar in two other experiments. (C) LLG-C4-GST (10 μg/ ml) was incubated in I domain–coated wells in the absence or presence of EDTA (2.5 mM), the LLG-C4 (1) peptide (100 μM), or the inactive LLG-C4(2) peptide (100 μM). The binding was determined with anti-GST antibodies. The results are the means ± SD from triplicate wells.

Article Snippet: The α IIb β 3 integrin antibody P2 was purchased from Immunotech, and the α V β 3 integrin antibody LM609 and the β 1 subunit antibody 6S6 were from Chemicon.

Techniques: Binding Assay, Purification, Control, Incubation

Journal: The Journal of Cell Biology

Article Title: Inhibition of β 2 Integrin–Mediated Leukocyte Cell Adhesion by Leucine–Leucine–Glycine Motif–Containing Peptides

doi:

Figure Lengend Snippet: Immobilized LLG-C4 supports β 2 integrin–directed cell adhesion. (A) Phorbol ester–activated THP-1 cells were allowed to bind for 60 min to microtiter wells coated with LLG-C4-GST, GST, or albumin. EDTA was included at a 2.5-mM concentration. The bound cells were determined by the assay measuring cellular phosphatase activity as described in Materials and Methods. The data are the means ± SD from triplicate wells. Similar results were obtained in six other experiments. (B) THP-1 cells were mixed with each antibody against the β 1 , β 2 , β 3 , α X , α M , or α L integrin subunit as indicated. An aliquot of cells was then transferred to wells coated with LLG-C4-GST and incubated for 60 min. The bound cells were determined by the phosphatase assay. The results are the mean percentage of adhesion ± SD of two to four independent experiments, each done in triplicate wells. (C) The C(1-8;3-9) and C(1-9;3-8) peptides were coated on microtiter wells in the absence or presence of glutaraldehyde. THP-1 cells (10 5 per well) were allowed to bind for 60 min and the bound cells were determined. The results show the mean ± SD of triplicate wells. The experiment was repeated twice. (D) The α X β 2 integrin–transfected L cells were allowed to bind to LLG-G4-GST or GST. The 7E4 antibody and EDTA were used as competitors. The results, mean percentage of adhesion ± SD, are representative of three experiments conducted in triplicate wells. The difference in the binding to LLG-C4-GST versus GST is statistically significant ( P = 0.016).

Article Snippet: The α IIb β 3 integrin antibody P2 was purchased from Immunotech, and the α V β 3 integrin antibody LM609 and the β 1 subunit antibody 6S6 were from Chemicon.

Techniques: Concentration Assay, Activity Assay, Incubation, Phosphatase Assay, Transfection, Binding Assay

Journal: The Journal of Cell Biology

Article Title: Inhibition of β 2 Integrin–Mediated Leukocyte Cell Adhesion by Leucine–Leucine–Glycine Motif–Containing Peptides

doi:

Figure Lengend Snippet: THP-1 cell adhesion to von Willebrand factor is inhibited by the LLG-C4 peptide. (A) THP-1 cell binding to von Willebrand factor was examined in the presence of antibodies against the β 2 (7E4), α V β 3 (LM609), or α IIb β 3 (P2) integrins. After a 60-min incubation in von Willebrand factor–coated wells, the bound cells were determined. The data are the mean ± SD of triplicate wells. The experiment was repeated three times. (B) THP-1 cell binding to wild-type von Willebrand factor or fibronectin in the presence of the indicated concentrations of C(1-8;3-9). (C) Binding of THP-1 cells to domain A2–deleted von Willebrand factor. The binding to wild-type von Willebrand factor was given as 100%. C(1-8;3-9) or RGD-4C was included as competitor at the concentrations described. The bound cells were determined by the phosphatase assay. The data are the mean ± SD of triplicate wells and were similar in two other experiments.

Article Snippet: The α IIb β 3 integrin antibody P2 was purchased from Immunotech, and the α V β 3 integrin antibody LM609 and the β 1 subunit antibody 6S6 were from Chemicon.

Techniques: Binding Assay, Incubation, Phosphatase Assay

Journal: Oncotarget

Article Title: Preliminary safety and imaging efficacy of the near-infrared fluorescent contrast agent DA364 during fluorescence-guided surgery in dogs with spontaneous superficial tumors

doi: 10.18632/oncotarget.27633

Figure Lengend Snippet: ( A ) Representative sigmoid curve revealing single-digit nanomolar affinity of DA364 for the human α v β 3 integrin receptor. ( B ) Dose-response inhibition of DA364 uptake in human melanoma WM266 cells by the unconjugated cRGD vector.

Article Snippet: A 96-well microtiter plate was coated overnight at 4° C with 1 μg/mL recombinant human α v β 3 integrin (Merck-Millipore, Darmstadt, Germany) in coating buffer (20 mM Tris HCl pH 7.4; 150 mM NaCl; 1 mM MnCl 2 ; 0.5 mM MgCl 2 ; 2 mM CaCl 2 ).

Techniques: Inhibition, Plasmid Preparation

Journal: Oncotarget

Article Title: Preliminary safety and imaging efficacy of the near-infrared fluorescent contrast agent DA364 during fluorescence-guided surgery in dogs with spontaneous superficial tumors

doi: 10.18632/oncotarget.27633

Figure Lengend Snippet: The workflow consists of the intravenous administration of DA364 24 h or 48 h prior to surgery ( A ), followed by pre-operative imaging ( B , C : procedure and fluorescent image in panel respectively), intra-operative imaging ( D , E : procedure and correspondent fluorescent image in panel respectively) and post-operative imaging ( F , G : procedure and correspondent fluorescent image, visualized on screen, respectively). Excised tissues were sliced in 4 μm-thick sections and underwent tumor assessment by Hematoxylin and Eosin histopathology ( H ), and evaluation of the β 3 integrin by immunohistochemistry ( I ).

Article Snippet: A 96-well microtiter plate was coated overnight at 4° C with 1 μg/mL recombinant human α v β 3 integrin (Merck-Millipore, Darmstadt, Germany) in coating buffer (20 mM Tris HCl pH 7.4; 150 mM NaCl; 1 mM MnCl 2 ; 0.5 mM MgCl 2 ; 2 mM CaCl 2 ).

Techniques: Imaging, Histopathology, Immunohistochemistry

Journal: Oncotarget

Article Title: Preliminary safety and imaging efficacy of the near-infrared fluorescent contrast agent DA364 during fluorescence-guided surgery in dogs with spontaneous superficial tumors

doi: 10.18632/oncotarget.27633

Figure Lengend Snippet: ( A ) Western blot analysis of integrin receptor subunits expression per tumor type. Black circles, malignant tumors; grey circles, benign tumors. Horizonal bars represent medians. ( B ) Representative image of β 3 integrin immunostaining on a mammary gland adenocarcinoma showing tumor cell and stromal staining (brown, β 3 integrin positive stain; cell nuclei are stained with Hematoxylin in blue). Scale bar, 100 microns.

Article Snippet: A 96-well microtiter plate was coated overnight at 4° C with 1 μg/mL recombinant human α v β 3 integrin (Merck-Millipore, Darmstadt, Germany) in coating buffer (20 mM Tris HCl pH 7.4; 150 mM NaCl; 1 mM MnCl 2 ; 0.5 mM MgCl 2 ; 2 mM CaCl 2 ).

Techniques: Western Blot, Expressing, Immunostaining, Staining

Journal: Oncotarget

Article Title: Preliminary safety and imaging efficacy of the near-infrared fluorescent contrast agent DA364 during fluorescence-guided surgery in dogs with spontaneous superficial tumors

doi: 10.18632/oncotarget.27633

Figure Lengend Snippet: Semi-quantitative (score) and quantitative (area) evaluation of anti-b 3 integrin staining in tumor samples

Article Snippet: A 96-well microtiter plate was coated overnight at 4° C with 1 μg/mL recombinant human α v β 3 integrin (Merck-Millipore, Darmstadt, Germany) in coating buffer (20 mM Tris HCl pH 7.4; 150 mM NaCl; 1 mM MnCl 2 ; 0.5 mM MgCl 2 ; 2 mM CaCl 2 ).

Techniques: Staining