We are currently varying two parameters of the system in order to find where the system operates best i.e. has the biggest difference in output between the +aBeta and -aBeta. The parameters are the amount of Receptor-TCS-Gal4VP16 construct transfected (since we have seen that a large amount of transfection places a strain on the cell) and how much TEVp-Cofilin is expressed.
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LilrB2
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Trial 1 Transfection
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PirB
Trial 1 Transfection Plan
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Trial 1 Transfection
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RESULTS
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Well 1
| Well 2hEF1a:tagBFP 200ng hEF1a:PirB 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Dummy 200ng | Well 3hEF1a:tagBFP 200ng hEF1a:PirB 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng | Well 4hEF1a:tagBFP 200ng hEF1a:PirB-TCS-G4VP16 200ng hEF1a:rtTA 200ng Gal4UAS:mKate 200ng Dummy 200ng | Well 5hEF1a:tagBFP 200ng hEF1a:PirB-TCS-G4VP16 200ng hEF1a:rtTA 200ng Gal4UAS:mKate 200ng Dummy 200ng 500 nM aBeta |
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| 100 ng PirB | |||
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Well 7hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 100ng Dummy 100 ng 1 nM Dox | Well 8hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 100ng Dummy 100 ng 1 nM Dox 500 nM aBeta | Well 9hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 100ng Dummy 100 ng 2000 nM Dox
| Well 10hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 100ng Dummy 100 ng 200 nM Dox 500 nM aBeta |
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| 50 ng PirB | |||
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Well 13hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 50ng Dummy 150 ng 10 nM Dox | Well 14hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 50ng Dummy 150 ng 10 nM Dox 500 nM aBeta | Well 15hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 50ng Dummy 150 ng 1000 nM Dox
| Well 16hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 50ng Dummy 150 ng 1000 nM Dox 500 nM aBeta |
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| 25 ng PirB | |
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Well 23hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 25 ng Dummy 175ng 100 nM Dox | Well 24hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 25 ng Dummy 175ng 100 nM Dox 500 nM aBeta |
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| 12.5 ng PirB | |||
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Well 19hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 12.5 ng Dummy 187.5 ng 10 nM Dox
| Well 20hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 12.5 ng Dummy 187.5 ng 10 nM Dox 500 nM aBeta | Well 21hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 12.5 ng Dummy 187.5 ng 1000 nM Dox
| Well 22hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 12.5 ng Dummy 187.5 ng 1000 nM Dox 500 nM aBeta |
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| 6.25 ng PirB | |||
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Well 11hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 6.25 ng Dummy 193.75 ng 1 nM Dox | Well 12hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 6.25 ng Dummy 193.75 ng 1 nM Dox 500 nM aBeta | Well 17hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 6.25 ng Dummy 193.75 ng 2000 nM Dox | Well 18hEF1a:tagBFP 200ng hEF1a:rtTA 200ng TRE:TEVp-Cofilin 200ng Gal4UAS:mKate 200ng hEF1a:PirB-TCS-Gal4VP16 6.25 ng Dummy 193.75 ng 2000 nM Dox 500 nM aBeta |
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