Description:
In this experiment, we want to test for the recruitment of our engineered cofilin to the activated receptor proteins. To do this, we will transfect HEK293 cells with LilrB2 and PirB as well as coflin-eYFP under a TRE promoter. We will then apply beta-amyloid oligomers in order to activate the receptors and add varying amounts of dox to induce varying levels of expression of cofilin-eYFP. We will look for increased yellow light intensity near the membrane of the cell. We test different levels of expression of cofilin because we don't know how the levels of endogenous cofilin in the cell will affect the interaction between our receptor and our engineered cofilin. We are looking for the level of expression of cofilin that will result in a discernible increased intensity of yellow near the membrane, indicating that a significant amount of cofilin-eYFP has recruited to the membrane.
* We can potentially test for the recruitment of cofilin to the receptors through co-immunoprecipitation.