These compounds may serve as the basis for the development of new treatments against Dengue virus infection. This procedure may be applied to other viral pathogens or for any other mechanism that involves specific conformational alterations for biological function. Our study also highlights the additional characteristics of certain tetracycline derivatives as effective inhibitors of DV propagation, which will allow further refinement of our screening program and potential medical application. Cell-based screens have been widely used in drug discovery although historically, these assays are conducted using genetically diverse cell lines derived from human tumors. Since the complex intracellular signaling networks that drive cancer cell growth and survival have begun to be elucidated, a more rational approach to drug discovery has become feasible. However, the implementation of target-orientated cell-based screens for anticancer drugs remains a challenge, both because of their reliance on defined genetic changes and because of the lack of proper control cells. To overcome this fundamental problem, we have developed a rational strategy for cell-based drug discovery that is based on the convenienceand flexibility of theBa/F3 cellsystem, animmortalized IL-3-dependent pro-B lymphoblastic cell line. IL-3 supports the growth and survival of Ba/F3 cells through the activation of distinct signaling pathways. Upon AZ 960 binding to its cognate receptor IL-3 activates the Janus kinase signal transduction and transcriptional activation pathways to induce Bcl-xL. Similarly, IL-3 activation of the PI3K/Akt pathway is involved in inhibiting the intrinsic apoptotic machinery in Ba/F3 cells. Overexpression of several SAR131675 constitutively active signaling molecules abrogates the dependence of these cells on IL-3. Hence, we generated isogenic cell lines derived from Ba/F3 in which IL-3 independent survival is sustained by independent signaling events. Each of these isogenic lines was genetically labeled with a fluorescent reporter and thus, the ratio of two spectrally distinct cell populations could be used as primary endpoint of the system to monitor pathway-specific cytotoxicity. Accordingly compounds can be screened in co-cultures of these lines and the change in the relative cell number of the two lines readily and rapidly measured to identify those molecules that specifically interact with one of the signaling pathways. In this instance, BaFiso has been designed as a live-cell system suitable to identify specific inhibitors of Akt signaling. The most frequently used anti-cancer therapies were discovered on the basis of their anti-proliferative activity in functional cell assays but with no pre-existing knowledge of the mechanism of action. As a result none of the current drugs directly targets the molecular lesions responsible for malignant transformation and they are not selective. Indeed this lack of selectivity between cancer cells and normal cells is currently one of the main reasons for the failure of conventional chemotherapy. In recent years, our understanding of the genetics of human cancer has increased rapidly, enabling more rational approaches to drug discovery for anti-cancer therapies to be adopted. Accordingly, the present study set out to develop a rational cell-based drug discovery strategy, an approach that has historically been compromised by the lack of appropriate control cells. With the objective of identifying lead compounds that specifically kill cells with activated Akt signaling and that spare control cells, we have combined the use of co-cultured isogenic cell lines with fluorescent technology. We introduced a myristoylated form of Akt which constitutively localizes to the plasma membrane, bypassing the requirement for PIP3 in Akt activation. This myr-Akt has been shown to constitutively inactivate proapoptotic downstream targets. In order to generate Ba/F3 cells that survive in the absence of IL-3 independent of activated PI3K/Akt signaling.