Integrin alpha V/CD51 Antibody NBP1-85746 from Novus Biologicals, a Bio-Techne Brand

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Cancer Res Commun
January 01 2023

Fig 1: Response of PDX UW-SCC22 to hSSTNIGF1R. A, UW-SCC22 PDXs implanted subcutaneously as bilateral tumors on the flanks of nude mice were allowed to establish for 1 week, followed by systemic delivery of 4.3 mg/kg/day hSSTNIGF1R via Alzet pump, saline alone delivered by Alzet pumps, or twice weekly subcutaneous injection of hSSTNIGF1R in saline. Tumor size (0.524 × length × width2) was documented for 4 weeks. B, Fixed, paraffin-embedded sections of tumors treated for 4 weeks with either saline or hSSTNIGF1R (pump delivery) are stained for expression of SDC1, IGF1R, or ITGAV compared with control goat (Gt) or rabbit (Rbt) IgG (bar = 50 µm). C, PLA is conducted between IGF1R and ITGAV in the tumors treated for 4 weeks either with saline, hSSTNIGF1R in Alzet pumps (pump) or tumors subjected to biweekly hSSTNIGF1R injection (inject) and is graphed as dots per cell (visualized using DAPI-stained nuclei). As controls, PLA is conducted using goat anti-IGF1R and nonspecific rabbit IgG or rabbit anti-ITGAV and nonspecific goat IgG (bar = 50 µm; ± SD, Student t test; ***, P = 0.001). D, Sections of tumors treated with saline or hSSTNIGF1R delivered by injection or pump for 4 weeks are stained with anti-pT845-ASK1 antibody compared with nonspecific IgG control (Ctrl; bar = 50 µm).
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Cancer Res Commun
January 01 2023

Fig 2: PLA screen of archival human head and neck carcinomas. A, Consecutive serial paraffin sections of eight archival human tongue tumors were stained for Sdc1, ITGAV, or IGF1R, or subjected to PLA to detect coupling of ITGAV and IGF1R. Shown are three of the six positive PLA cases (cases #2–4) representative of the range of observed staining, including the two tumor cases containing adjacent benign tissue, along with the two cases (cases #9 and #1) demonstrating negative PLA. Tissue histology as viewed by hematoxylin and eosin (H&E) staining and benign tissue and the tumor margins are marked (dashed line), if present. Triangles within the benign tissue (B) or tumor (T) identify the areas examined by PLA at higher magnification (bar = 50 µm). B, Quantification of ITGAV:IGF1R PLA intensity (PLA dots/cell) in the eight cases of tongue carcinoma arranged in order of intensity (PLA is quantified as dots per cell across 15 images over three experiments). Case #5 was excluded because of high endogenous fluorescence. C, Quantification of ITGAV:IGF1R PLA conducted on three tissue microarrays [one adenoid cystic carcinoma microarray and two oropharyngeal squamous cell carcinoma (OPSCC) microarrays]. PLA quantified as fluorescent dots per cell for each tissue based on the averaged staining of three microarray sections and expressed as a violin plot (solid line is the median and dotted lines are quartiles). Range for the tumors is 0.000–17.704 dots per cell and benign tissue is 0.004–0.056. Negative control staining of the same sections ranges from 0.000 to 0.057 dots per cell.
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Cancer Res Commun
January 01 2023

Fig 3: Detection of assembled receptor complex using proximity ligation assay. A, Staining of fixed UM-SCC47 cells pretreated for 6 hours with vehicle or 30 µmol/L hSSTNIGF1R, then stained for SDC1, IGF1R, or ITGAV compared with control rabbit (Rbt) or goat (Gt) IgG (bar = 50 µm). Data are representative of duplicate experiments. B, Model of PLA between ITGAV and IGF1R. C, PLA between ITGAV and IGF1R (ITGAV:IGF1R) in fixed UM-SCC47 cells pretreated for 6 hours with vehicle (left top) or 30 µmol/L hSSTNIGF1R (right top), compared with PLA between ITGAV and control goat IgG or IGF1R and control rabbit IgG in vehicle-treated cells (bottom left; bar = 50 µm). Individual cells are identified by DAPI-stained nuclei. PLA is quantified as dots per cell across 15 images over three experiments and graphed as a percentage of PLA observed in untreated cells for ease of comparison (± SD, Student t test; ***, P = 0.001). For reference, the average for the untreated cells was 72 dots per cell.