1. Inhibition of Experimental Lung Metastasis by Aerosol Delivery of PEI–p53 Complexes 
Ajay Gautam, Charles L. Densmore, J.Clifford Waldrep 
Molecular Therapy 
Volume 2, Issue 4, Pages (October 2000)
DOI: /mthe
Copyright © 2000 American Society for Gene Therapy Terms and Conditions

2. FIG. 1
Effect of p53 on proliferation of B16-F10 cells in vitro. (a) One hundred thousand B16-F10 cells/well were plated on a 12-well plate and incubated for 24 h at 37°C. The monolayer was then transfected with PEI, PEI–Luc, or PEI–p53 complexes for 24 h at a PEI:DNA ratio of 5:1 with 2 μg/ml DNA concentration. The cell number was counted at different time points to determine the cell proliferation. (b) Twenty thousand B16-F10 cells/well were plated on a 48-well plate. After incubating for 24 h at 37°C, the cultures were transfected with PEI–Luc or PEI–p53 for 24 h at a fixed PEI:DNA ratio of 5:1 with increasing concentrations of DNA (0.5, 1, 2, and 4 μg/ml). The cell proliferation after 24 h was determined by the Alamar blue assay. The values for 0.5, 1, and 2 μg/ml DNA concentration between Luc and p53 are statistically significant (P < 0.5), but not for 4 μg/ml (P >
Molecular Therapy 2000 2, DOI: ( /mthe )
Copyright © 2000 American Society for Gene Therapy Terms and Conditions

3. FIG. 2
Inhibition of B16-F10 lung metastasis by PEI–p53 aerosol delivery. Mice were injected via lateral tail vein with 25,000 B16-F10 cells on day 0. The mice were then treated with PEI alone, PEI–Luc, or PEI–p53 aerosol complexes twice a week starting on day 1. The last treatment was on day 22 post tumor cells injection, making it a total of seven aerosol exposures. The mice were sacrificed on day 24, the lungs harvested, weighed and fixed in Fekets solution. (a) Tumor index was calculated by the formula: Tumor index = lung weights × average grade for the group. Values are means ± SD (n = 10 mice per group). (b) Representative lungs from control, PEI–Luc and PEI–p53-treated mice are presented (n = 10 mice per group). Lungs from PEI-treated group (not shown) were similar in shape, size, and number of tumor foci to those shown for control and PEI–Luc-treated groups. Data are representative of two separate experiments. (c) Lung weights of mice from different groups. Values are means ± SD (n = 10 mice per group).
Molecular Therapy 2000 2, DOI: ( /mthe )
Copyright © 2000 American Society for Gene Therapy Terms and Conditions

4. FIG. 3
Survival time of mice treated with PEI–p53 aerosol complexes. Mice (n = 10 mice per group) were injected intravenously with 25,000 B16-F10 cells on day 0. The mice were then exposed to PEI, PEI–Luc, or PEI–p53 aerosol complexes on days 1, 4, 8, and 11. The mice were monitored over time for their survival. Each step represents the number of mice dead at the indicated time point. Data are representative of two separat
Molecular Therapy 2000 2, DOI: ( /mthe )
Copyright © 2000 American Society for Gene Therapy Terms and Conditions