Vascular endothelial growth factor expression in breast cancer cell lines and in circulating tumor cells. A: VEGF, VEGFR2 HIF-1α and pFAK expression in cultured breast cancer cell lines, 1: MCF7, 2:T47D, 3: SKBR3 4:MDA MB-453, 5: MDA MB-231 (western blot analysis of equal protein amount of cell lysates). B (I): Representative images of confocal laser scanning microscopy of PBMCs cytospins stained with pancytokeratins A45 B/B3 (green), and VEGF anti-goat (red) antibodies. B (II): Representative images of PBMCs cytospins stained with pancytokeratins A45 B/B3 (green), VEGF anti-goat (red) antibodies and Dapi (blue). B (III): Quantification of VEGF and CK co-expression in CTCs of 34 metastatic breast cancer patients. All the examined patients (100%) presented CTCs in their blood, while 62% also presented double positive cells (VEGF+CK+). (C) Quantification of double positive CTCs/total CTCs for each examined molecule in 34 metastatic breast cancer patients. For pFAK testing cytospins from 12 patients were evaluated. indicatively. Seventy-three percent of the examined CTCs were VEGF+CK+ positive, 71% were VEGFR2+CK+, 56% were HIF-1α+CK+ and 81% were pFAK+CK+.