Relative mRNA expression of melanoma genes as a function of time after i.v. tumor inoculation.

<p>(A) Mice were injected i.v. with 10⁵ B16.F10gp melanoma cells and lungs were collected 2, 4, 8, 24, 96 hours or 11 days later and snap frozen in liquid nitrogen. Total RNA was extracted and converted into cDNA and the expression of GP100 (white) and TRP-2 (grey) was detected using triplex Q-PCR. The experiment was performed twice with similar outcomes. (B) Mice were infected i.v. with 10⁶ pfu VSV or left untreated. One day later, depleting anti-NK1.1 antibodies were administered i.p. to some groups and on the next day, all mice were injected i.v. with 10⁵ B16.F10gp cells. Lungs were collected 4 hours after tumor inoculation and the relative mRNA expression of GP100 (white) and TRP-2 (grey) was obtained using triplex Q-PCR analysis. The relative mRNA expression in the lungs of one mouse collected 2 hours after tumor inoculation, and of one mouse vaccinated with Ad-Ii-OVA and lungs isolated 6 days after tumor challenge (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087831#pone-0087831-g005" target="_blank">Fig. 5B</a>) were used as calibrators in A and B, respectively, and set to 1. Shown are medians and ranges of 5 mice per group; results are representative of two experiments. WT = wild type, IFNγ = interferon gamma deficient, Pfp = perforin deficient, * denotes p<0.05.