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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: Oral Oncol. 2011 Oct 19;48(1):26–32. doi: 10.1016/j.oraloncology.2011.09.010

Figure 4.

Figure 4

Endothelial secreted VEGF induces phosphorylation of Akt in HNCSC. (A–D) Western blots depicting the expression of total and phosphorylated Akt in ALDH+CD44+ or ALDH−CD44− cells sorted from UM-SCC-22A or UM-SCC-22B. Cells were cultured in serum-free conditioned medium collected from primary endothelial cells (EC CM) supplemented with 1 μg/ml anti-VEGF antibody or non-specific isotype matched IgG control in ultra-low attachment plates. As additional controls, we evaluated cells cultured in presence of EC CM without IgG for 60 minutes, or in presence of serum-free endothelial basal medium (EBM) supplemented (or not) with 50 ng/ml rhVEGF165 during all time points. (E) Western blots depicting the expression of VEGFR1 and VEGFR2 in unsorted UM-SCC-22A and UM-SCC-22B cells, using primary human endothelial cells (HDMEC) as controls. (F) Western blot to evaluate the effect of EC CM on the expression of VEGFR1 in ALDH+CD44+ UM-SCC-22A and UM-SCC-22B cells, using unsorted HDMEC as controls.