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Testing effectiveness of miRNA1–miRNA4 in down-regulating BACE1 expression v1

A1

    • Hef1a: eBFP 250ng
    • Dummy DNA 750ng


      Positive Control

A2

    • Hef1a: eBFP 250ng
    • Hef1a: BACE1 250ng
    • Dummy DNA 500ng

Negative Control

A3

    • Hef1a: eBFP 250ng
    • TRE: miRNA1-mKATE250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      0 nM Dox

A4

    • Hef1a: eBFP 250ng
    • TRE: miRNA1-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

0 nM Dox

A5

    • Hef1a: eBFP 250ng
    • TRE: miRNA1-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      1000 nM Dox

A6

    • Hef1a: eBFP 250ng
    • TRE: miRNA1-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

1000 nM Dox

A7

  • Hef1a:mKATE 250ng
  • Hef1a:BFP 250ng
  • Dummy DNA 500ng
     
      
      

Testing inducible expression of BACE2

B1

  • Hef1a:eBFP 333ng
  • Dummy DNA 666ng

Negative Control

B2

  • Hef1a:eBFP 333ng
  • Hef1a:rtTA 333ng
  • TRE:BACE2 333ng

0 nM Dox

B3

  • Hef1a:eBFP 333ng
  • Hef1a:rtTA 333ng
  • TRE:BACE2 333ng

1000 nM Dox

B4

  • No DNA (untransfected)

 

Double negative control

  
      
      
      

Transfection: Week of 8/11

A1

    • Hef1a: eBFP 250ng
    • Dummy DNA 750ng



Negative BACE1 Control

A2

    • Hef1a: eBFP 250ng
    • Hef1a: BACE1 250ng
    • Dummy DNA 500ng

 

Positive BACE1 Control

 A3

  • Hef1a:mKATE 250ng
  • Hef1a:BFP 250ng
  • Dummy DNA 500ng

mKATE control

A4

  • Hef1:miRNAg1 250ng
  • Hef1a:BFP 250ng
  • Dummy DNA 500ng

miRNA1g control

  
      
      

    • Hef1a:mKATE 250ng
    • Hef1a:BFP 250ng
    • Dummy DNA 500ng

 

mKATE control

A4

    • Hef1:miRNAg1 250ng
    • Hef1a:BFP 250ng
    • Dummy DNA 500ng

 

miRNA1g control

A5

    • Hef1a: eBFP 250ng
    • TRE: miRNA1-mKATE250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      0 nM Dox

A6

    • Hef1a: eBFP 250ng
    • TRE: miRNA1-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

0 nM Dox

A7

    • Hef1a: eBFP 250ng
    • TRE: miRNA1-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      1000 nM Dox

A8

    • Hef1a: eBFP 250ng
    • TRE: miRNA1-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

1000 nM Dox

A9

    • Hef1a: eBFP 250ng
    • TRE: miRNA2-mKATE250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      0 nM Dox

A10

    • Hef1a: eBFP 250ng
    • TRE: miRNA2-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

0 nM Dox

A11

    • Hef1a: eBFP 250ng
    • TRE: miRNA2-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      1000 nM Dox

A12

    • Hef1a: eBFP 250ng
    • TRE: miRNA2-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

1000 nM Dox

A13

    • Hef1a: eBFP 250ng
    • TRE: miRNA3-mKATE250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      0 nM Dox

A14

    • Hef1a: eBFP 250ng
    • TRE: miRNA3-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

0 nM Dox

A15

    • Hef1a: eBFP 250ng
    • TRE: miRNA3-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      1000 nM Dox

A16

    • Hef1a: eBFP 250ng
    • TRE: miRNA3-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

1000 nM Dox

A17

    • Hef1a: eBFP 250ng
    • TRE: miRNA4-mKATE250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      0 nM Dox

A18

    • Hef1a: eBFP 250ng
    • TRE: miRNA4-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

0 nM Dox

A19

    • Hef1a: eBFP 250ng
    • TRE: miRNA4-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Dummy DNA 250ng

      1000 nM Dox

A20

    • Hef1a: eBFP 250ng
    • TRE: miRNA4-mKATE 250ng
    • Hef1a: rtTA 250ng
    • Hef1a: BACE1 250ng

1000 nM Dox

  

B1

    • Hef1a:eBFP 333ng
    • Dummy DNA 666ng

 

Negative Control

B2

    • Hef1a:eBFP 333ng
    • Hef1a:rtTA 333ng
    • TRE:BACE2 333ng

 

0 nM Dox

B3

    • Hef1a:eBFP 333ng
    • Hef1a:rtTA 333ng
    • TRE:BACE2 333ng

 

1000 nM Dox

B4

    • No DNA (untransfected)

 

 

Double negative control