Description:
In this description, we are trying to asses the extent of the cleavage of the TCS fused to the receptors by TEVp-Cofilin given different levels of expression of TEVp-Cofilin. Cleavage of the TCS will result in the release of the transcription factor Gal4VP16 which will in turn activate the Gal4UAS driven mKate.
We have reason to believe that there are high levels of endogenous cofilin expression in HEK293 cells (cofilin is used as a loading control fro Western blots, lol). The levels of endogenous vs exogenous cofilin will be assessed through the cofilin Western blot experiment. However, if this is the case, we don't expect to see any mKate Gal4UAS activation at low levels of expression of TEVp-Cofilin as the endogenous cofilin is likely to overwhelm the exogenous TEVp-Cofilin. Hopefully, we do get an increase in mKate expression at some high level of dox induction. This may indicate that we could just express TEVp-Coflin constitutively.
LilrB2:
PirB:
Trial 1
We are doing the first PirB Trial without using beta-amyloid to activate the receptor, since we are not actually sure whether or not it binds the receptor.
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Controls: Well 2: Bleed through from eBFP into mKate Well 3: Leaky expression of Gal4UAS:mKate without the presence of Gal4VP16 Well 4: Background activation of Gal4UAS:mKate without presence of TEVp-Cofilin
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Trial 2
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Controls: Well 2: Bleed through from eBFP into mKate Well 3: Leaky expression of Gal4UAS:mKate without the presence of Gal4VP16 Well 4: Background activation of Gal4UAS:mKate without presence of TEVp-Cofilin Well 5: Positive control
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Results
LilrB2
PirB
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