Fig. 2

Fig. 2

a Schematic of Lewis Lung Carcinoma (LLC) tumor allograft assay with representative tumor images. Bar = 1 cm. b–gCds2 vMO treatment (n = 10 biologically independent tumors for all groups). h–l L-690,488 small molecule inhibitor treatment (control n = 8 biologically independent tumors, L-690,488 n = 19 biologically independent tumors). Quantitation of average tumor volume (b, h), final tumor weight (c, i) and final tumor vascular density (d, j) at 12–15 days post-tumor implantation, in control versus Cds2 vMO or L-690,488-treated animals (b–d versus h–j). d Quantitation of average tumor vascular density and representative images of CD31/PECAM labeled (green) versus DAPI (blue) LLC tumor sections from control vMO (top) and Cds2 vMO (bottom) treated mice. White arrowheads indicate sites of CD31/PECAM positive blood vessel labeling. Bar = 100 μm. For all vascular density measurement experiments: three images per tumor were acquired and vascular density measured for all groups. A minimum of two slide sections from each tumor in b–g (taken from sections at least 10 slices apart) were analyzed. Representative of three experimental replicates. e-g PIP2 “rescues” Cds2 vMO tumor inhibition. Quantitation of LLC average tumor volume (e), final tumor weight (f), and tumor vascular density (g) at 18 days post-tumor implantation in control vMO (n=8 biologically independent tumors), Cds2 vMO #2 (no liposomes, n = 9 biologically independent tumors), Cds2 vMO #2 + carrier liposome (no PIP2, n = 9 biologically independent tumors), or Cds2 vMO #2 + PIP2 loaded liposome-treated (n = 8 biologically independent tumors), LLC-allografted mice. Data in (e-g) are normalized to the average starting size of each individual tumor group at day 4, and shown as a percentage of the starting day 4 control (the PIP2 liposome injection start date). j Quantitation of average tumor vascular density of LLC tumor sections from control and L-690,488-treated mice. For all vascular density measurement experiments: three images per tumor were acquired and vascular density measured for all groups. A minimum of two slide sections from each tumor in h–l (taken from sections at least 10 slices apart) were analyzed. Representative of two experimental replicates. k, l Myo-inositol “rescues” L-690,488 tumor inhibition. Quantitation of LLC average tumor volume (k) and final tumor weight (l) at 15 days post-tumor implantation in control (untreated, n = 14 biologically independent tumors), myo-inositol (n = 11 biologically independent tumors), L-690,488 (n = 12 biologically independent tumors), or L-690,488 + myo-inositol (n = 10 biologically independent tumors) treated LLC-allografted mice. Data in (k, l) are normalized to the control condition. For all panels: p ≤ 0.05, error bars ± SEM. Star indicates significance from control (t-test). 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.