1. Date of download: 5/27/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Molecular Targeting of Ultrasonographic Contrast Agent for Detection of Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2012;138(7):662-668. doi:10.1001/archoto.2012.1081 Figure 1. Bar graft of flow cytometry analysis. Four head and neck cancer cell lines (SCC-1, SCC-5, FaDu, and Cal27), 2 CD147 knock-down cell lines (SCC-1/SiE-711 and SCC-5/SiE-711), and normal dermal fibroblasts (NDFs) were analyzed by flow cytometry to assess cell surface expression of epidermal growth factor receptor (EGFR) and extracellular matrix-metalloproteinase inhibitor (CD147). FITC indicates fluorescein isothiocyanate. Error bars indicate SE. ** P <.01. *** P <.001. Figure Legend:

2. Date of download: 5/27/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Molecular Targeting of Ultrasonographic Contrast Agent for Detection of Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2012;138(7):662-668. doi:10.1001/archoto.2012.1081 Figure 2. Microbubble (MB) binding to receptor expression. Correlation of MB EGFR (A), MB CD147 (B), and MB DUAL (C) binding to receptor expression was assessed in 4 head and neck cancer cell lines (SCC-1, SCC-5, FaDu, and Cal27), 2 head and neck cancer CD147 knock-down cell lines (SCC-1/SiE-711 and SCC-5/SiE-711), and normal dermal fibroblasts (NDFs). FITC indicates fluorescein isothiocyanate. MB IgG, MB EGFR, MB CD147, and MB DUAL indicate IgG-, anti-EGFR–, and anti-CD147–targeted MBs and dual-conjugated MBs, respectively. Figure Legend:

3. Date of download: 5/27/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Molecular Targeting of Ultrasonographic Contrast Agent for Detection of Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2012;138(7):662-668. doi:10.1001/archoto.2012.1081 Figure 3. Dark-field microscopy images and statistical comparison. A, Dark-field microscopy images (original magnification ×200) of MB IgG, MB EGFR, MB CD147, and MB DUAL binding to SCC-1, FaDu, and normal dermal fibroblast (NDF) cells. B, Microbubble binding per cell was calculated, and statistical comparison was made between agents. MB IgG, MB EGFR, MB CD147, and MB DUAL indicate IgG-, anti- EGFR–, and anti-CD147–targeted MBs and dual-conjugated MBs, respectively. * P <.05. ** P <.01. *** P <.001. Figure Legend:

4. Date of download: 5/27/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Molecular Targeting of Ultrasonographic Contrast Agent for Detection of Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2012;138(7):662-668. doi:10.1001/archoto.2012.1081 Figure 4. Representative ultrasonographic (US) images of SCC-1 flank tumors after injection with MB IgG, MB EGFR, MB CD147, and MB DUAL. A, White dashed ovals represent targeted contrast agent in tumor microvasculature. B, Images were analyzed using ImageJ, and average intratumoral echogenicity was compared between targeted agents. Error bars indicate SE. C, Linear correlation of tumor size through caliper and high-frequency US measurement. AU indicates arbitrary units. MB IgG, MB EGFR, MB CD147, and MB DUAL indicate IgG-, anti-EGFR–, and anti-CD147–targeted MBs and dual-conjugated MBs, respectively. Figure Legend:

5. Date of download: 5/27/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Molecular Targeting of Ultrasonographic Contrast Agent for Detection of Head and Neck Squamous Cell Carcinoma Arch Otolaryngol Head Neck Surg. 2012;138(7):662-668. doi:10.1001/archoto.2012.1081 Figure 5. Immunohistochemistry staining of FaDu and SCC-1 flank tumors in athymic nude mice. Hematoxylin-eosin staining show tumor cells and anti-CD147 and anti-EGFR immunostaining show ubiquitous expression of head and neck squamous cell carcinoma used to confirm tumor pathology (original magnification ×200). Figure Legend: