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Table 1 Depletion of FANCA by RNA interference does not directly lead to Increased ATM gene expression as assessed by Q-PCR.

From: Upregulated ATM Gene Expression and Activated DNA Crosslink-Induced Damage Response Checkpoint in Fanconi Anemia: Implications for Carcinogenesis

  

Transient

Stable

  

siRNA 1

siRNA 3

Control

siRNA 1

siRNA 3

Control

H-CYPB

      
 

Average

21.11

21.31

20.91

20.87

20.53

19.52

 

SD

0.03

0.00

0.01

0.00

0.05

0.17

H-FANCA

      
 

Average

27.11

27.60

25.86

26.96

26.80

24.61

 

SD

0.00

0.19

0.02

0.15

0.02

0.17

 

Delta Ct

6.00

6.29

4.95

6.09

6.27

5.09

 

SD

0.03

0.19

0.02

0.15

0.06

0.24

 

Test-control

1.05

1.34

0.00

1.00

1.18

0.00

 

Fold change

0.48

0.40

1.00

0.50

0.44

1.00

H-ATM

      
 

Average

25.89

26.71

25.04

24.83

24.96

22.87

 

SD

0.05

0.00

0.00

0.15

0.08

0.14

 

Delta Ct

4.78

5.40

4.13

3.95

4.44

3.36

 

SD

0.06

0.01

0.01

0.15

0.10

0.22

 

Test-control

0.65

1.27

0.00

0.59

1.08

0.00

 

Fold change

0.64

0.41

1.00

0.66

0.47

1.00

  1. Either transient (left columns) or stable (right columns) transfection of shRNA against FANCA was performed: FANCA shRNA sequences 1, 3, and empty vector control, respectively. Knockdown of FANCA mRNA expression by either shRNA construct was confirmed when compared with the empty vector control (assigned a value of 1.0). Transient or stable depletion of FANCA mRNA expression led to modestly decreased levels of ATM gene expression. All comparisons were made using human cyclophilin B (H-CYPB) as an internal control. Shown are: Average cycle threshold; SD; delta Ct (difference between Ct of test and the H-CYPB internal control); test — control (difference between delta Ct of test and the empty vector control); fold change (1/2Δ−ΔCt).
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Molecular Medicine

ISSN: 1528-3658