Figure 2. In vivo Matrigel angiogenesis induced by mouse and human breast tumor cells with Ncoa1 knockout and NCOA1 knockdown, respectively.

a. Representative images of Matrigel plugs with angiogenesis induced by W1, W2, K1 and K2 cells in SCID/beige mice (upper four panels) and representative images of CD31 and Flk1 immunofluorescent staining (green color) and DAPI staining (blue color) of Matrigel plug sections with angiogenesis induced by W1 and K1 cells (lower four panels). b. Representative images of Matrigel plugs with angiogenesis induced by K1 and K2 cells as well as 10 nM of recombinant VEGFa (rVEGFa) protein in Matrigel (upper two panels) and representative images of CD31 and Flk1 immunofluorescent staining and DAPI staining of Matrigel plug sections with angiogenesis induced by K1 cells plus rVEGFa (lower two panels). c. Semi-quantitative analysis of angiogenesis induced by W1, W2, K1 and K2 cells as well as K1 and K2 cells plus rVEGFa in Matrigel plugs in mice. A total of 12 Matrigel plugs with angiogenesis induced by the indicated cells with or without rVEGFa were analyzed. The number of microvessels per 200× viewing field was counted and 5 viewing fields were examined for each Matrigel plug. The data are presented as Mean ± SD. *p < 0.05 by Student's t test. d. Representative images of Matrigel plugs with angiogenesis induced by MDA-MB-231 cells expressing non-targeting shRNA (shCtrl) or NCOA1 mRNA-targeting shRNAs (sh1 and sh2) and representative images of CD31 and Flk1 immunofluorescent staining and DAPI staining of Matrigel plug sections with angiogenesis induced by MDA-MB-231 cells expressing shCtrl or sh1. e. Semi-quantitative analysis of angiogenesis induced by MDA-MB-231 cells expressing shCtrl, sh1 or sh2 in Matrigel plugs. Microvessels in 6 Matrigel plugs for each group were examined and counted as described above. Data are presented as Mean ± SD. *p < 0.05 by Student's t test. The knockdown efficiency of NCOA1 was analyzed by Western blot.