Figure 2.

Macroscopic view of whole lungs and hematoxylin and eosin (H&E)-stained lung sections from Pep 2-treated mice. (A) Representative images of lungs surgically removed from healthy animals or from animals intravenously injected with HT1080 cells: of these, 5 were untreated (vehicle), 5 were treated with Pep 2 either at 15 mg/kg or at 3.4 mg/kg. (B) H&E-stained of the corresponding representative lung sections with the neoplastic areas marked with T (4× magnification, Scale bar 100 µm. (C) 50 ng of total genomic DNA extracted from lung samples from each vehicle, or Pep 2-treated mice were used as template for semi-quantitative PCR reactions and compared to the healthy lungs (H). The amplification products are separated on 1% agarose gels. A standard curve was included in every run, generated by mixing 10⁷, 10⁶,10⁵,10⁴,10³,10²,10¹,1 HT1080 human cells with 1 to 10⁷ murine P19 cells (Standard 1 to 9). The uncropped image for agarose gel can be found in Figure S5. (D) The densitometric analysis of PCR amplification products, accomplished by acquiring images and quantifying them through ImageJ software (NIH, Bethesda, MD, USA), was performed on three independent gels. The signal from the 1 × 10⁷ HT1080 cells sample was considered as 100% and standards 2, 3, 4 and 5 were graded relative to that (** p < 0.005; *** p < 0.001, Student’s t-test).