Page History

Versions Compared

Old Version 6

changes.mady.by.user Unknown User (lylaatta@mit.edu)

Saved on Oct 02, 2014 20:08

compared with

New Version 7

changes.mady.by.user Unknown User (lylaatta@mit.edu)

Saved on Oct 02, 2014 20:26

Key

This line was added.
This line was removed.
Formatting was changed.

Comment: Reverted from v. 5

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

RESULTS

Controls

Well 1

Well 2

hEF1a:tagBFP 200ng

hEF1a:

PirB

LilrB2 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Dummy 200ng

Well 3

hEF1a:tagBFP 200ng

hEF1a:

PirB

LilrB2 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

Well 4

hEF1a:tagBFP 200ng

hEF1a:

PirB

LilrB2-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

Well 5

hEF1a:tagBFP 200ng

hEF1a:

PirB100 ng PirB

LilrB2-TCS-G4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate 200ng

Dummy 200ng

500 nM aBeta

Image Removed

Image Removed

Image Removed

Image Removed

Image Removed

Well 6

Well 7

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:

PirB

LilrB2-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

LilrB2-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

LilrB2-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

LilrB2-TCS-Gal4VP16 100ng

Dummy 100 ng

200 nM Dox

500 nM aBeta

Image Removed

Image Removed

Image Removed

Image Removed50 ng PirB

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

1 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

1 nM Dox

500 nM aBeta

Well 13

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:

PirB

LilrB2-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

LilrB2-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

LilrB2-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

LilrB2-TCS-Gal4VP16 50ng

Dummy 150 ng

1000 nM Dox

500 nM aBeta

Image Removed

Image Removed

Image Removed25 ng PirB

Well 23

Image Removed

Well 17

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:

PirB

LilrB2-TCS-Gal4VP16 6.25 ng

Dummy

175

193.75 ng

100

2000 nM Dox

Well

24

18

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:

PirB

LilrB2-TCS-Gal4VP16 6.25 ng

Dummy

175

193.75 ng

100

2000 nM Dox

500 nM aBeta

Image Removed

Image Removed12.5 ng PirB

Well 19

hEF1a:tagBFP 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

hEF1a:

PirB

LilrB2-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

LilrB2-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

LilrB2-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

LilrB2-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:LilrB2-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:LilrB2-TCS-Gal4VP16 25 ng

Dummy 175ng

100 nM Dox

500 nM aBeta

Image Removed

Image Removed

Image Removed

Image Removed 6.25 ng PirB

PirB

Trial 1 Transfection Plan

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

Image Removed

Image Removed

Image Removed

Image Removed

Image Removed

Image Removed

Image Removed

Image Removed

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