Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Association of the α 2 δ 1 Subunit with Ca v 3.2 Enhances Membrane Expression and Regulates Mechanically Induced ATP Release in MLO-Y4 Osteocytes

doi: 10.1002/jbmr.437

Figure Lengend Snippet: RT-PCR primers used for detection of VSCC subunits and osteocyte markers

Article Snippet: The rabbit anti-β 3 antibody was purchased from Alomone Research Laboratories (Jerusalem, Israel).

Techniques: Sequencing

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