Group E: Testing TEVp-Cofilin cleavage with active receptors

Created by Unknown User (lylaatta@mit.edu), last modified on Sep 15, 2014 11:20

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Description:

In this experiment, we are testing whether or not beta amyloid activates the receptors, recruiting TEVp-Cofilin and leading to higher TCS cleavage and in turn, higher Gal4UAS activation. From experiment D, we determined that the level of TEVp-Cofilin expression corresponding to 0 nM dox was the level at which there was the least background TCS cleavage. We are, therefore, using TRE:TEVp-Cofilin with no dox induction in our experiment.

LilrB2:

PirB:

Well 1

HEK293

Well 2

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin200ng

Dummy 200ng

Well 3

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin200ng

Gal4UAS:mKate200ng

Well 4

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate200ng

Dummy 200ng

Well 4

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

Gal4UAS:mKate200ng

Dummy 200ng

500 nM a beta

Well 7

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

0nM DOX

Well 8

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

1nM DOX

Well 9

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

10nM DOX

Well 10

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

1000nM DOX

Well 11

Well 12

Well 13

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

0nM DOX

500 nM aBeta

Well 14

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

1nM DOX

500 nM

Well 15

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

10nM DOX

500 nM

Well 16

HEK293

hEF1a:eBFP 200ng

hEF1a:PirB-TCS-Gal4VP16 200ng

hEF1a:rtTA 200ng

TRE:TEVp-Cofilin 200ng

Gal4UAS:mKate 200ng

10000nM DOX

500 nM

Well 17

Well 18

Well 19

Well 20

Well 21

Well 22

Well 23

Well 24

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