In vitro and in vivo expression of AdNK4.
(A) Detection of AdNK4-mediated NK4 expression by RT-PCR in B16-F10 cells transduced with AdNK4. The generated cDNA from B16-F10 cells transduced with AdNK4, AdNull, or PBS alone at a multiplicity of infection of 100 for 3 days was amplified with primers for NK4 or GAPDH mRNA. (B) Detection of AdNK4-mediated NK4 expression by RT-PCR in B16-F10 tumors transduced with AdNK4. To demonstrate the expression of AdNK4 in vivo, C57Bl/6 mice were injected subcutaneously in the right flank with 3 × 105 B16-F10 cells (day 0). On day 8, tumor-bearing mice were treated by intratumoral injection of 109 pfu of AdNK4, AdNull, or PBS alone. On day 11, the generated cDNA from the transduced B16-F10 tumors was amplified with primers for NK4 or GAPDH mRNA. For both panels, PCR products were resolved in a 1% agarose gel and stained with ethidium bromide.
