| Well 1 | Well 2 | Well 3 | Well 4 | Well 5 | Well 6 | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
untransfected (calibration) | eBFP bleed through control | leaky mKate control | TEVp control (nonspecific G4 cleavage) | TEVp control (nonspecific G4 cleavage) | TEVp control (nonspecific G4 cleavage) | ||||||
HEK293
| HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B TRE: Syk-TEVp | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B TRE: Syk-TEVp UAS: mKate
| HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B UAS: mKate | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B-TCS-G4 UAS: mKate | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B-TCS-G4 UAS: mKate | ||||||
| Well 7 | Well 8 | Well 9 | Well 10 | Well 11 | Well 12 | Well 2 HEK 293
UAS: mKate | Well 3 HEK293
| Well 4 HEK293 | Well 5 HEK293 | Well 6 HEK293 | Well 7|
HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B TRE: Syk-TEVp UAS:mKate 0nM dox Well 8 | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B TRE: Syk-TEVp UAS:mKate 1nM dox | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B TRE: Syk-TEVp UAS:mKate 10nM doxWell 9 | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B TRE: Syk-TEVp UAS:mKate 10nM dox 100nM dox | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B TRE: Syk-TEVp UAS:mKate 1000nM doxWell 10 | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B TRE: Syk-TEVp UAS:mKate 100nM dox Well 112000nM dox | ||||||
| Well 13 | Well 14 | Well 15 | Well 16 | Well 17 | Well 18 | ||||||
HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 0nM dox | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 1000nM dox 1nM dox | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 10nM dox | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 100nM doxWell 12 | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 1000nM dox | HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 2000nM dox
| ||||||
| Well 19 | Well 20 | Well 21 | Well 22 | Well | 1323 | Well 24 | |||||
HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 0nM dox
| Well 14 HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 1nM dox
| Well 15 HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 10nM dox
| Well 16 HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 100nM dox
| Well 17 HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 1000nM dox
| Well 18 HEK293 hEF1a: eBFP hEF1a: rtTA GmabH GmabL CD79A-TCS-G4 CD79B-TCS-G4 TRE: Syk-TEVp UAS:mKate 2000nM dox |
| Untransfected | eBFP + Syk-TEVp + BCR (no Gal4VP16) | eBFP + Syk-TEVp + BCR (no Gal4VP16) + UAS:mKate | eBFP + BCR (CD79A-TCS-G4, CD79B) + UAS:mKate | eBFP + BCR (CD79A, CD79B-TCS-G4) + UAS:mKate | eBFP + BCR (CD79A-TCS-G4, CD79B-TCS-G4) + UAS:mKate | |
|---|---|---|---|---|---|---|
 |
Transfection Efficiency: ~20% |  |  |  |  | |
| eBFP + BCR (CD79A-TCS-G4, CD79B) + UAS:mKate | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B) + UAS:mKate [0nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B) + UAS:mKate [1nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B) + UAS:mKate [10nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B) + UAS:mKate [100nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B) + UAS:mKate [1000nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B) + UAS:mKate [2000nM dox] |
|
Cotransfection Efficiency: ~4% |  |  |  |  |  |
| eBFP + BCR (CD79A, CD79B-TCS-G4) + UAS:mKate | eBFP + Syk-TEVp + BCR (CD79A, CD79B-TCS-G4) + UAS:mKate [0nM dox] | eBFP + Syk-TEVp + BCR (CD79A, CD79B-TCS-G4) + UAS:mKate [1nM dox] | eBFP + Syk-TEVp + BCR (CD79A, CD79B-TCS-G4) + UAS:mKate [10nM dox] | eBFP + Syk-TEVp + BCR (CD79A, CD79B-TCS-G4) + UAS:mKate [100nM dox] | eBFP + Syk-TEVp + BCR (CD79A, CD79B-TCS-G4) + UAS:mKate [1000nM dox] | eBFP + Syk-TEVp + BCR (CD79A, CD79B-TCS-G4) + UAS:mKate [2000nM dox] |
|
Cotransfection Efficiency: ~3.3% |  |  |  |  |  |
| eBFP + BCR (CD79A-TCS-G4, CD79B-TCS-G4) + UAS:mKate | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B-TCS-G4) + UAS:mKate [0nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B-TCS-G4) + UAS:mKate [1nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B-TCS-G4) + UAS:mKate [10nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B-TCS-G4) + UAS:mKate 100nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B-TCS-G4) + UAS:mKate [1000nM dox] | eBFP + Syk-TEVp + BCR (CD79A-TCS-G4, CD79B-TCS-G4) + UAS:mKate [2000nM dox] |
|
Cotransfection efficiency: ~3.3% |  |  |  |  |  |
