T, or GFPMCF7 CXCR4CTD tumors at three, 4, and five wk just after orthotopic implantation of cells into either the third or the fourth mammary gland. Encircled areas represent regions of interest for assessment of tumor development. (c) Representative photos of key tumors and lymph node metastases of GFPMCF7 CXCR4WT tumors. Fluorescence microscopy of metastases from a tumor bearing GFPMCF7 CXCR4WT cells in the inguinal lymph node near the tumor from mouse 1 (xenograft inside the fourth mammary gland) and the axillary lymph nodes close to the tumor from mice 3 (xenograft inside the third mammary gland). Tissues from tumor and lymph nodes were dissected and examined working with fluorescence microscopy. Bars, 150 m. (d) Representative images of main tumors and lymph node metastases of GFPMCF7 CXCR4CTD tumors. Fluorescence microscopy of metastases from a tumor bearing GFPMCF7 CXCR4CTD cells at the inguinal lymph node near the tumor from mice 1 and two (xenograft inside the fourth mammary gland) plus the axillary lymph nodes near the tumor from mice 3 and 4 (xenograft in the third mammary gland). 576 | T. Sobolik et al.Molecular Biology of your CellFIGURE 7: Behavior of GFP MCF7 CXCR4WT and GFP MCF7 CXCR4CTD cells in vivo. (a ) Intravital pictures from a time series of GFPMCF7 vector, GFPMCF7CXCR4WT, and GFPMCF7 CXCR4CTD cells orthotopically implanted within the absence of exogenous estrogen in the fourth mammary fat pad of athymic nude mice two wk before imaging. Host vasculature was labeled with 30 l of 20 mg/ml rhodamine dextran (70 kDa), a skin flap was made to expose the mammary fat pad, and pictures have been acquired with an LSM 510 META inverted confocal microscope having a 200.75 Strategy Apochromat objective. (a) GFPMCF7 vector tumors have been not detected in mice in absence of exogenous estrogen. The GFP and Texas red channels are shown. The trajectories (yellow) of myeloid cells (red) tracked for 20 min with Bitplane Imaris are shown (Supplemental Motion pictures S1 and S2). (b, c) MCF7 CXCR4WT cells migrate toward blood vessels in singlecell streams. The mostdisplaced trajectories (yellow) of single GFPMCF7 CXCR4WT cells (green) and myeloid cells (red) in the tumor tracked for 20 min are shown (Supplemental Motion pictures S3 and S4). (d) GFPMCF7 CXCR4WT are nonmigratory, and GFPMCF7 CXCR4CTD cells show random migration in tumors with out a vasculature. The mostdisplaced trajectories (yellow) of single GFPMCF7 CXCR4WT cells (green) and GFPMCF7 CXCR4CTD cells (green) in the tumor tracked for 20 min are shown (Supplemental Motion pictures S5 and S6).Volume 25 March 1, 2014 The function of CXCR4 in breast cancer|FIGURE eight: MCF7 CXCR4CTD cells migrate toward blood vessels and metastasize towards the lymph nodes.[Ir(dF(Me)ppy)2(dtbbpy)]PF6 Formula (a) Intravital images from a time series of GFPMCF7 CXCR4CTD cells orthotopically implanted in the fourth mammary fat pad of athymic nude mice 2 wk prior to imaging.4-Chloro-6-methyl-7-azaindole site Differentiated HL60 cells had been labeled with DiI Cy5 (blue) and injected in to the vasculature through a catheter within the femoral vein.PMID:23805407 Host vasculature was labeled with 30 l of 20 mg/ml rhodamine dextran (70 kDa), a skin flap was created to expose the mammary fat pad, and pictures were acquired two h just after injection of labeled HL60 cells with an LSM 510 META inverted confocal microscope with a 401.3 Strategy Apochromat objective. An asterisk is placed over the area of reference to act as a landmark to determine the path of cell movement. Differentiated HL60 cells (average of two cells within the vasculature adjacent for the tumor) labeled with DiI Cy5 (blue) are indicated by.