Table 4

Differential expression of selected SSH transcripts by quantitative real-time polymerase chain reaction (QRT-PCR) ***

Genes

Normalized CT values

log2 CT X1000000

Fold Change (Delta CT Values)*


Average (n = 3)

SEM


LD

LM

CD

CM

LD

LM

CD

CM

LD

LM

CD

CM

CD/CM

CM/CD

LD/LM

LD/CD

LM/CM


Cts h

26.5

24.2

26.1

24.1

0.03

0.01

0.05

0.01

96

19

70

18

3.9

0.3

5.0

1.4

1.1

Eif4ebp2

36.5

30.4

33.7

30.9

0.62

0.07

0.07

0.05

95252

1430

13833

1974

7.0

0.1

66.6

6.9

0.7

Gst m1

23.4

19.1

23.2

18.2

0.32

0.02

0.00

0.06

11

1

9

0

29.9

0.0

19.7

1.2

1.8

Gst o1

36.0

26.9

35.5

29.8

0.14

0.17

0.56

0.06

68660

125

47023

909

51.7

0.0

550.9

1.5

0.1

Serpina3n

21.8

21.1

26.1

23.5

0.03

0.02

0.09

0.05

4

2

72

12

6.1

0.2

1.6

0.1

0.2

Sprr 2A

29.4

21.8

25.4

19.8

0.06

0.14

0.03

0.00

697

4

45

1

51.0

0.0

185.1

15.3

4.2

Upk1a

32.5

24.4

30.0

23.7

0.18

0.01

0.02

0.13

6162

23

1050

14

76.4

0.0

271.4

5.9

1.7

Upk1b

26.5

22.2

26.7

21.4

0.04

0.02

0.12

0.05

98

5

112

3

39.3

0.0

20.6

0.9

1.7

Calm2

23.2

24.0

22.2

23.8

0.11

0.09

0.04

0.11

9

17

5

14

0.3

3.0

0.6

2.0

1.2

Cnn1

32.1

33.1

27.2

31.0

0.50

0.09

0.15

0.05

4547

9400

149

2177

0.1

14.6

0.5

30.5

4.3

Smoc2

33.9

34.7

26.0

28.4

0.06

0.27

0.04

0.06

15953

27402

68

352

0.2

5.2

0.6

234.8

77.7

Bcap31

33.3

29.2

26.7

26.9

0.40

0.07

0.08

0.01

10331

627

106

127

0.8

1.2

16.5

97.6

4.9

Catenin

36.9

33.0

27.1

28.0

0.23

0.07

0.42

0.09

130085

8592

143

273

0.5

1.9

15.1

912.8

31.5

Mafb

32.1

30.1

31.8

30.5

0.65

0.10

0.34

0.44

4703

1154

3615

1570

2.3

0.4

4.1

1.3

0.7

Pls3

37.4

35.3

28.6

29.3

0.10

0.18

0.14

0.15

179012

41906

402

661

0.6

1.6

4.3

444.9

63.4

Prss11

29.9

27.9

25.8

24.5

0.05

0.03

0.06

0.00

999

259

59

23

2.5

0.4

3.9

17.1

11.0

Syn1

30.1

25.3

24.1

22.8

0.22

0.04

0.21

0.13

1148

42

18

7

2.4

0.4

27.3

65.6

5.9

Lamp2

31.6

31.1

28.1

27.8

0.21

0.14

0.05

0.01

3342

2379

285

230

1.2

0.8

1.4

11.7

10.3

Mpp1

31.0

30.0

29.2

29.5

0.44

0.11

0.11

0.19

2114

1090

632

753

0.8

1.2

1.9

3.3

1.4

Sepp1

23.0

23.9

22.0

21.6

0.06

0.03

0.08

0.05

8

15

4

3

1.4

0.7

0.6

2.0

4.8

Thsd6

28.1

28.3

28.3

28.0

0.13

0.08

0.03

0.04

293

329

338

272

1.2

0.8

0.9

0.9

1.2

IFIT3

30.4

31.2

30.8

30.9

0.08

0.00

0.12

0.05

1455

2396

1885

1988

0.9

1.1

0.6

0.8

1.2

Ifrg15

27.9

27.9

28.2

27.6

0.03

0.05

0.09

0.07

254

245

307

210

1.5

0.7

1.0

0.8

1.2

Sdfr1

24.4

25.6

24.1

24.8

0.06

0.09

0.10

0.09

23

52

18

29

0.6

1.6

0.4

1.3

1.8

Siva

26.7

25.8

27.1

25.8

0.03

0.07

0.06

0.07

110

59

147

59

2.5

0.4

1.9

0.7

1.0

Prnp

24.1

25.7

23.3

24.6

0.02

0.07

0.02

0.00

18

53

11

26

0.4

2.5

0.3

1.7

2.0


*(ratio of antilog2 of cycle threshold values)

** (bolded cells indicate values greater than 3.0)

***Female C57BL/6J mice were instilled with saline (n = 20) or LPS (n = 20). Twenty four hours after LPS instillation, mice were euthanized, the bladder was removed and placed in RNAlaterâ„¢ (Ambion) for separation of the mucosa and submucosa from the detrusor smooth muscle, as described in Material and Methods. Four sample groups were obtained as follows: control mucosa (CM), control detrusor (CD), LPS-treated mucosa (LM), and LPS-treated detrusor (LD). The QRT-PCR amplifications were accomplished on an ABIPRISM 7700 using SYBRGreen I dye assay chemistry

All samples were run in triplicate with the appropriate single QRT-PCR controls (no reverse transcriptase and no template). From the QRT-PCR data, an average cycle threshold (Ct) value was calculated from the triplicate reactions. Averaged Ct values were then normalized (to adjust for different amounts of cDNA within each reaction) to the exongenous control gene, RCA. The relative expression level of each transcript within each sample group (CD, CM, LD, and LM) was determined by calculating the ratio of the antilog2 of the delta Ct values.

Saban et al. BMC Physiology 2006 6:1   doi:10.1186/1472-6793-6-1

Open Data