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Reactivation of Src signaling in NECTIN4-inhibited angiosarcoma cells. Control or NECTIN4 siRNA-transfected HAMON and ISO-HAS-B cells were further treated with <t>MLR-1023</t> and assessed for cell proliferation and angiogenesis. ( A,B ) Mean (± SD) number of viable cells in siRNA-transfected, DMSO (0.1%) or MLR-1023 (1 μM)-treated ( A ) HAMON and ( B ) ISO-HAS-B cells, as detected by the CCK-8 assay. Data show fold changes relative to Day 0. Experiments were repeated three times, with three wells used for each condition. Significance of differences was calculated between NECTIN4-siRNA transfected DMSO-treated cells and NECTIN4-siRNA transfected MLR-1023-treated cells. ( C,D ) Representative images of angiogenesis in ( C ) HAMON and ( D ) ISO-HAS-B cells. Scale bars = 0.5 mm. ( E,F ) Number of tubules per 1 cm 2 , tubule length per 1 cm 2 , and the number of junctions per 1 cm 2 of ( E ) HAMON and ( F ) ISO-HAS-B cells in the angiogenesis assay. Data are the mean ± SD of three independent experiments. ** P < 0.01, and *** P < 0.001.
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Reactivation of Src signaling in NECTIN4-inhibited angiosarcoma cells. Control or NECTIN4 siRNA-transfected HAMON and ISO-HAS-B cells were further treated with MLR-1023 and assessed for cell proliferation and angiogenesis. ( A,B ) Mean (± SD) number of viable cells in siRNA-transfected, DMSO (0.1%) or MLR-1023 (1 μM)-treated ( A ) HAMON and ( B ) ISO-HAS-B cells, as detected by the CCK-8 assay. Data show fold changes relative to Day 0. Experiments were repeated three times, with three wells used for each condition. Significance of differences was calculated between NECTIN4-siRNA transfected DMSO-treated cells and NECTIN4-siRNA transfected MLR-1023-treated cells. ( C,D ) Representative images of angiogenesis in ( C ) HAMON and ( D ) ISO-HAS-B cells. Scale bars = 0.5 mm. ( E,F ) Number of tubules per 1 cm 2 , tubule length per 1 cm 2 , and the number of junctions per 1 cm 2 of ( E ) HAMON and ( F ) ISO-HAS-B cells in the angiogenesis assay. Data are the mean ± SD of three independent experiments. ** P < 0.01, and *** P < 0.001.

Journal: Scientific Reports

Article Title: Nectin cell adhesion molecule 4 regulates angiogenesis through Src signaling and serves as a novel therapeutic target in angiosarcoma

doi: 10.1038/s41598-022-07727-x

Figure Lengend Snippet: Reactivation of Src signaling in NECTIN4-inhibited angiosarcoma cells. Control or NECTIN4 siRNA-transfected HAMON and ISO-HAS-B cells were further treated with MLR-1023 and assessed for cell proliferation and angiogenesis. ( A,B ) Mean (± SD) number of viable cells in siRNA-transfected, DMSO (0.1%) or MLR-1023 (1 μM)-treated ( A ) HAMON and ( B ) ISO-HAS-B cells, as detected by the CCK-8 assay. Data show fold changes relative to Day 0. Experiments were repeated three times, with three wells used for each condition. Significance of differences was calculated between NECTIN4-siRNA transfected DMSO-treated cells and NECTIN4-siRNA transfected MLR-1023-treated cells. ( C,D ) Representative images of angiogenesis in ( C ) HAMON and ( D ) ISO-HAS-B cells. Scale bars = 0.5 mm. ( E,F ) Number of tubules per 1 cm 2 , tubule length per 1 cm 2 , and the number of junctions per 1 cm 2 of ( E ) HAMON and ( F ) ISO-HAS-B cells in the angiogenesis assay. Data are the mean ± SD of three independent experiments. ** P < 0.01, and *** P < 0.001.

Article Snippet: MMAE (ChemScene, Deerpark, NJ; CS-0837), LY294002 (Abcam; ab120243), dasatinib (Abcam; ab142050), MLR-1023 (Sigma-Aldrich, St. Louis, MO; SML0361), and cabozantinib (Selleck Biotech, Tokyo, Japan; BMS-907351) were dissolved in DMSO (Sigma-Aldrich; 07-4860-5), which was also used as the vehicle control (final concentration 0.1% DMSO).

Techniques: Transfection, CCK-8 Assay, Angiogenesis Assay