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Fig. 1 | Breast Cancer Research

Fig. 1

From: Polyfunctional anti-human epidermal growth factor receptor 3 (anti-HER3) antibodies induced by HER3 vaccines have multiple mechanisms of antitumor activity against therapy resistant and triple negative breast cancers

Fig. 1

Human epidermal growth factor receptor 3 (HER3)-specific antibody responses are induced by adenovirus encoding full length human HER3 (Ad-HER3) in vivo. a Binding of vaccine-induced antibody (VIA) in serum from mice immunized with Ad-HER3 (HER3-VIA), Ad-lacZ (lacZ-VIA), and Ad-green fluorescence protein (GFP-VIA). Serum at dilutions presented were mixed with the HER3-4 T1 cell line or wild-type 4 T1 and binding of antibody was identified with near infrared (nIR) dye-conjugated anti-mouse IgG and detected by a LI-COR Odyssey Imager. The difference in fluorescence intensity between HER3-4 T1 and 4 T1 is graphed. b Binding of HER3-VIA to breast cancer cell lines: a panel of human breast cancer cell lines were incubated with dilutions of the HER3-VIA, washed, and then mixed with a phycoerythrin (PE)-conjugated secondary antibody. HER3-VIA binding was analyzed based on fluorescence activated cell sorting analysis and mean fluorescence intensity was reported. c Flow cytometric analysis was used to identify percentage of BT474 cells able to bind HER3-VIA (1:100 dilution). d Binding of His-tagged heregulin to HER3 receptor was measured by pre-incubating BT474 cells with heregulin (0, 10, 100 nM), or HER3-VIA (1:100) for 10 min on ice, followed by incubation with His-tagged heregulin (100 nM) for 10 min. Receptor-bound His-tagged heregulin was visualized by staining with PE-conjugated anti-His Tag antibody. Mean fluorescence intensity is shown in each histogram

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