Fig. 5

Fig. 5

a Schematic of established tumor allograft assay. Lewis Lung Carcinoma (LLC) tumor cells were injected into each flank of adult B6/C57 mice at day 0 and tumors allowed to develop for 12–15 days. Daily intravenous injections of control vivoMorpholino (vMO) or vMO targeting one of two independent sites in the Cds2 gene (vMO #1 and vMO #2) were started at day 4 after tumor implantation. Volume measurements were taken daily starting at day 4, when tumors became visible and vMO treatment started, with final tumor volumes and weights taken at the termination of the experiment. b Quantitation of daily average tumor volume (in mm³) of LLC tumors, normalized to the average starting size of each tumor condition at day 4 (the injection start date). c Quantitation of average tumor weight (in mg) of LLC tumors at 14 days post-tumor implantation. Control vMO (n = 9); Cds2 vMO #1 (n = 8); Cds2 vMO #2 (n = 6). d Images of tumors collected from control vMO or Cds2 vMO-treated animals at 14 days post-tumor implantation. Bar = 1 cm. e Quantitation of LLC tumor vessel density in control vMO- versus Cds2 vMO-treated mice. f–h Representative images of CD31/PECAM labeled (green) LLC tumor sections from control vMO (f) and Cds2 vMO (g, h) treated mice. White arrowheads indicate sites of CD31/PECAM positive blood vessel labeling. For all vascular density measurement experiments, three images per tumor were acquired (see tumor numbers above) and vascular density measured for all groups. A minimum of two slide sections from each tumor (taken from sections at least 10 slices apart) were utilized for independent analysis. Bar = 100 μm. Box plots are graphed showing the median versus the first and third quartiles of the data (the middle, top, and bottom lines of the box, respectively). The whiskers demonstrate the spread of data within 1.5x above and below the interquartile range. All data points are shown as individual dots, with outliers shown above or below the whiskers.