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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Mol Cancer Ther. 2014 Jun 4;13(8):2050–2061. doi: 10.1158/1535-7163.MCT-13-1063

Figure 5.

Figure 5

HEY β5 integrin knockdown impairs soft agar and tumor growth with reduced FAK Y397 phosphorylation in spheroids. A, stable HEY β5 integrin knockdown by lentiviral shRNAs (β5-1 and β5-2) as determined by immunoblotting with actin as a loading control. B, flow cytometry of cell surface αvβ5 levels in Scr, β5-1 and β5-2 HEY cells. Black histogram is secondary antibody only. C, mean adherent HEY growth over 4 days (+/− SD, * p<0.05). D, Mean Scr, β5-1 and β5-2 HEY soft agar colony growth over 7 days (+/− SD, * p<0.05, *** p<0.001). E, representative spheroid fluorescent staining for pY397 FAK (red), OPN (green), and cell nuclei (blue) of Scr and β5-1 shRNA HEY cells. Scale is 50 μm. F, orthotopic tumor growth of Scr (n=7), β5-1 (n=9), and β5-2 (n=10) HEY cells in the ovarian bursa. Values are mean tumor mass after 21 days (+/− SD, * p<0.05, ** p<0.01). G, serum from Scr and β5-1 HEY tumor-bearing mice was analyzed by anti-hOPN immunoblotting. Densitometry was used to determine mean values (n=7 tumors each, +/− SD, * p<0.05).