FIG. 1.

Schematic of dual fluorescence reporter mouse model. (A) Lentiviral constructs used to generate the dual reporter cell line. vTD147 expresses enhanced green fluorescent protein (EGFP) under the control of the human phosphoglycerate kinase (PGK) promoter. vTD171 expresses tdTFP containing the mRNA sequence from click beetle luciferase (CBL) that is targeted by CBL3 siRNA in the 3′ untranslated region (UTR) under the control of the human ubiquitin C (UbC) promoter. (B) Schematic representation of the reporter mouse model in which red fluorescent protein (tdTFP) is targeted by siRNA treatment. Human A375 melanoma cells are transduced with lentiviral constructs expressing vTD147 and vTD171 to generate a dual reporter cell line. Following injection of labeled melanoma cells into immunocompromised mice, the resulting tumors are treated with siRNA. Successful delivery of functional siRNA results in specific inhibition of tdTFP. EGFP should not be affected by siRNA treatment and is therefore used as a normalization control.