Titration of ligands for the stimulation of MDA-MB-231 BRE and CAGA reporter cell lines

Considerable amount of ligands will be consumed in the dual luciferase assay (DLA) which will be used for the screening of ALK2 inhibitors and off-target inhibition of ALK5 routinely. Ligands are costly to procure. In order to minimise wastage, it is important to determine the least amount of ligands needed to achieve optimal stimulation of ALK2 and ALK5. In these experiments, the activation of ALK2 and ALK5 by different concentrations of ligands in MDA-MB-231 reporter cells was analysed in Western Blot and DLA.