Live Cell Imaging of the TGF- β/Smad3 Signaling Pathway In Vitro and In Vivo Using an Adenovirus Reporter System

This method can hep answer key questions in the cell signaling field by noninvasively tracking cell signaling in real time. The main advantage of this technique is that adenovirus systems provide a quick and easy method for cell signaling detection that can achieve a 100%infection rate in cancer cell lines. The implications of this technique extend toward therapeutic development for cancers and can be expanded to other signaling pathways.

Though this method can provide insight into breast cancer models, it can also be applied to other cancer types, such as brain, bone, and lung. On the day of adenovirus infection, seed the wells of a 12-well plate with approximately 1.25 times 10 to the fifth MDA-MB-231 cells in 500 microliters of medium for a confluency of 35%Infect cells in all wells with 75 microliters of adenovirus-CMV-GFP and 7.5 microliters of adenovirus CAGA12-Td-Tomato to obtain a 2, 500 MOI which can achieve 100%infection positivity. Then add TGF-beta to the treatment well immediately after adenovirus infection.