Fig. 3

Intratumoral FoxP3 mRNA transcripts before and after treatment with and without denileukin diftitox. Fresh biopsies was collected before and after treatment when feasible and snap frozen in liquid nitrogen. RNA samples were isolated using the Trizol regent, and cDNA was synthesized using the High Fidelity cDNA Synthesis Kit (Roche Applied Science). qRT-PCR was then performed to analyze the FoxP3 transcripts levels on ABI-7300 qPCR instrument (Applied Biosystem). β-actin were used as an internal control