Initial Screening of CD79B-X-TCS-X-Gal4VP16 was largely uninformititve. The data did not appear to be statistically significant. We think this is likely due to minimal expression of the B-Cell Receptor ~5% drowning out our signal. To attempt to boost our signal, we are optimizing receptor expression relative to Syk-X-TEV/tTEV. Although our data appeared to be insignificant, we selected our three most promising candidates based on normalizing the fluorescent signal to our transfection marker. The linkers selected are S12 12TCS12, S9 9TCS15, and S6 12TCS6.
| Untransfected | Single Color Blue | Background anti-IgM binding | Single Color Red | Double Color | -VE control (background activation) | -VE control (background cleavage) | +VE control |
|---|---|---|---|---|---|---|---|
| HEK293 | HEK293 hEF1a:MabH hEF1a:MabL hEF1a:Lyn
hEF1a:CD79A-15TCS15-G4VP16 hEF1a:CD79B TRE:Syk-15-tTEVp Dox x uM | HEK293 hEF1a:MabH hEF1a:MabL hEF1a:Lyn
hEF1a:CD79A-15TCS15-G4VP16 hEF1a:CD79B TRE:Syk-15-tTEVp Dox x uM anti-IgM | HEK293 hEF1a:MabH hEF1a:MabL hEF1a:Lyn
hEF1a:CD79A-15TCS15-G4VP16 hEF1a:CD79B TRE:Syk-15-tTEVp Dox x uM | HEK293 hEF1a:MabH hEF1a:MabL hEF1a:Lyn hEF1a:CD79A-15TCS15-G4VP16 hEF1a:CD79B TRE:Syk-15-tTEVp Dox x uM | HEK293 hEF1a:MabH hEF1a:MabL hEF1a:Lyn UAS:mKate hEF1a:CD79A
hEF1a:CD79B TRE:Syk-15-tTEVp Dox x uM | HEK293 hEF1a:MabH hEF1a:MabL hEF1a:Lyn UAS:mKate hEF1a:CD79A-15TCS15-G4VP16
hEF1a:CD79B TRE:Syk Dox x uM | HEK293 hEF1a:MabH hEF1a:MabL hEF1a:Lyn UAS:mKate hEF1a:CD79A
hEF1a:CD79B TRE:Syk-15-tTEVp hEF1a:GAl4VP16 Dox x uM |
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