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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.

Expand

title	Parameter Selection

Gliffy Diagram

name	Tuning

LilrB2

Expand

title	LilrB2 Trial 1 Transfection Plan

Trial 1 Transfection

Well 1

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

Well 2

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

500 nM aBeta

Well 3

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

2000 nM Dox

Well 4

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

200 nM Dox

500 nM aBeta

Well 5

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

1 nM Dox

Well 6

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

1 nM Dox

500 nM aBeta

Well 7

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

Well 8

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

500 nM aBeta

Well 9

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

Well 10

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

500 nM aBeta

Well 11

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

2000 nM Dox

Well 12

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

2000 nM Dox

500 nM aBeta

Well 13

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

10 nM Dox

Well 14

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

10 nM Dox

500 nM aBeta

Well 15

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

1000 nM Dox

Well 16

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

1000 nM Dox

500 nM aBeta

Well 17

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

Well 18

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

500 nM aBeta

Well 19

HEK 293

Well 20

hEF1a:tagBFP 200ng

hEF1a:LilrB2 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Dummy 200ng

Well 21

hEF1a:tagBFP 200ng

hEF1a:LilrB2 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

Well 22

hEF1a:tagBFP 200ng

hEF1a:LilrB2-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

Well 23

hEF1a:tagBFP 200ng

hEF1a:LilrB2-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

500 nM aBeta

Well 24

PirB

Expand

title	PirB Trial 1 Transfection Plan

Trial 1 Transfection

Well 1

Well 2

hEF1a:tagBFP 200ng

hEF1a:PirB 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Dummy 200ng

Well 3

hEF1a:tagBFP 200ng

hEF1a:PirB 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

Well 4

hEF1a:tagBFP 200ng

hEF1a:PirB-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

Well 5

hEF1a:tagBFP 200ng

hEF1a:PirB-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

500 nM aBeta

Well 6

Well 7

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

Well 8

hEF1a: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 9

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 100ng

Dummy 100 ng

2000 nM Dox

Well 10

hEF1a: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

Well 11

hEF1a: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 12

hEF1a: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 13

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

Well 14

hEF1a: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 15

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

Well 16

hEF1a: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

Well 17

hEF1a: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 18

hEF1a: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

Well 19

hEF1a: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 20

hEF1a: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 21

hEF1a: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 22

hEF1a: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

Well 23

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

Well 24

hEF1a: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

Expand

title	PirB Trial 2 Transfection Plan

Well 1

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

Well 2

hEF1a: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 3

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 100ng

Dummy 100 ng

2000 nM Dox

Well 4

hEF1a: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

Well 5

hEF1a: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 6

hEF1a: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 7

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

Well 8

hEF1a: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 9

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

Well 10

hEF1a: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

Well 11

hEF1a: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 12

hEF1a: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

Well 13

hEF1a: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 14

hEF1a: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 15

hEF1a: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 16

hEF1a: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

Well 17

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

Well 18

hEF1a: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

Well 19

Well 20

hEF1a:tagBFP 200ng

hEF1a:PirB 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Dummy 200ng

Well 21

hEF1a:tagBFP 200ng

hEF1a:PirB 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

Well 22

hEF1a:tagBFP 200ng

hEF1a:PirB-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

Well 23

hEF1a:tagBFP 200ng

hEF1a:PirB-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

500 nM aBeta

Well 24

RESULTS

LilrB2

Expand

title	PirB LilrB2 Trial 1

Well 2

Controls

Well 19

HEK 293

Well 20

hEF1a:tagBFP 200ng

hEF1a:LilrB2 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Dummy 200ng

Well 21

hEF1a:tagBFP 200ng

hEF1a:LilrB2 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

Well 1

Well 22

hEF1a:tagBFP 200ng

hEF1a:LilrB2-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

Well 23

hEF1a:tagBFP 200ng

hEF1a:LilrB2-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

500 nM aBeta

Image Added

Image Added

Image Added

Image Added

Image Added

100 ng LilrB2

Well 1

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

Well 2

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

500 nM aBeta

Well 3

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

2000 nM Dox

Well 4

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

200 nM Dox

500 nM aBeta

Image Added

Image Added

Image Added

Image Added

50 ng LilrB2

Well 7

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

Well 8

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

500 nM aBeta

Well 9

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

Well 10

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

500 nM aBeta

Image Added

Image Added

Image Added

Image Added

25 ng LilrB2

Well 17

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

Well 18

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

500 nM aBeta

Image Added

Image Added

12.5 ng LilrB2

Well 13

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

10 nM Dox

Well 14

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

10 nM Dox

500 nM aBeta

Well 15

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

1000 nM Dox

Well 16

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

1000 nM Dox

500 nM aBeta

6.25 ng LilrB2

Well 5

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

1 nM Dox

Well 6

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

1 nM Dox

500 nM aBeta

Well 11

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

2000 nM Dox

Well 12

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:LilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

2000 nM Dox

500 nM aBeta

PirB

Expand

title	PirB Trial 1

Controls

Well 1

Well 2

hEF1a:tagBFP 200ng

hEF1a:PirB 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Dummy 200ng

Well 3

hEF1a:tagBFP 200ng

hEF1a:PirB 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

Well 4

hEF1a:tagBFP 200ng

hEF1a:PirB-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

Well 5

hEF1a: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

Well 7

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

Well 8

hEF1a: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 9

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 100ng

Dummy 100 ng

2000 nM Dox

Well 10

hEF1a: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

Well 13

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

Well 14

hEF1a: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 15

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

Well 16

hEF1a: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

Well 23

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirB-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

Well 24

hEF1a: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

Well 19

hEF1a: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 20

hEF1a: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 21

hEF1a: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 22

hEF1a: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

Well 11

hEF1a: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 12

hEF1a: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 17

hEF1a: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 18

hEF1a: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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Expand

title	PirB Trial 2

Controls

Well 19

HEK 293

Well 20

hEF1a:tagBFP 200ng

hEF1a:LilrB2 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Dummy 200ng

Well

3

21

hEF1a:tagBFP 200ng

hEF1a:PirB LilrB2 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

Well

4

22

hEF1a:tagBFP 200ng

hEF1a:PirBLilrB2-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

Well

5

23

hEF1a:tagBFP 200ng

hEF1a:PirBLilrB2-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

500 nM aBeta

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100 ng PirBLilrB2

Well

7

1

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

Well

8

2

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

1 nM Dox

500 nM aBeta

Well

9

3

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

2000 nM Dox

Well

10

4

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

200 nM Dox

500 nM aBeta

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50 ng PirBLilrB2

Well

13

7

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

Well

14

8

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

10 nM Dox

500 nM aBeta

Well

15

9

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

Well

16

10

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

500 nM aBeta

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25 ng PirBLilrB2

Well

23

17

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

Well

24

18

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

500 nM aBeta

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12.5 ng PirBLilrB2

Well

19

13

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

10 nM Dox

Well

20

14

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

10 nM Dox

500 nM aBeta

Well

21

15

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

1000 nM Dox

Well

22

16

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 12.5 ng

Dummy 187.5 ng

1000 nM Dox

500 nM aBeta

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6.25 ng PirBLilrB2

Well

11

5

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

1 nM Dox

Well

12

6

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

1 nM Dox

500 nM aBeta

Well

17

11

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

2000 nM Dox

Well

18

12

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:PirBLilrB2-TCS-Gal4VP16 6.25 ng

Dummy 193.75 ng

2000 nM Dox

500 nM aBeta

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