Supplementary Figure Legends
Supplementary Figure 1: Detection of human IGF-IR mRNA in the liver following HD-CBsIGFIR injection. Total RNA was extract from frozen liver fragments 72 hrs following virus inoculation as described in the legend to Fig 3 and analyzed by RT-PCR using the primers (forward): 5’-TTGCCCGAAGGTCTGTGAG-3’ and (reverse) 5’-GGAGCAGTAATTGTGCCGG-3’ that amplify a fragment of 1025 bp specific to human IGF-IR. The GAPDH transcript was used as an internal control and amplified using the primers (forward) 5’-GGTGAAGGTCGGTGTGAACGGATTT-3’ and (reverse) 5’-AATGCCAAAGTTGTCATGGATGACC-3’. cDNA amplification conditions were as previously described (1) and the DNA fragments analyzed by electrophoresis on 1% agarose gels without further purification. Mouse colon carcinoma MC-38 and human colon carcinoma KM12SM cells that express high levels of the respective IGF-IR (2) were used as controls for primer specificity. Shown is a representative gel of 2 separate analyses.
Supplementary Figure 2: Lack of virus neutralizing activity in mice sera following a single injection of virus particles. Virus neutralizing activity was analyzed using a fluorescence-based assay (3). Sera collected from mice 1, 7 and 21 days following the injection of 1-4X1011 (days 1 and 21) or 1.2X1012 (day 7) HD-CBsIGFIR particles were heat inactivated, serially diluted and AdV HCA-GFPCBLacZ particles (4) were added at a MOI of 250. The mixture was added to 104 A549 cells in a 96-well plate for a 24 hr incubation at 370C. The A549 cells were fixed using 2% formaldehyde and fluorescence was measured at an excitation wave length of 488nm a using a plate reader. Results are expressed as fold change (±SD) relative to normal (uninfected) mouse serum used at the same dilution and are based on sera obtained from 3 different mice.
* -p<0.1 as analyzed by the student’s t-test.
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