Table 4 |
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|
Differential expression of selected SSH transcripts by quantitative real-time polymerase chain reaction (QRT-PCR) *** |
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|
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 |
|
|
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|
*(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 RNAlater™ (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. |
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|
Saban et al. BMC Physiology 2006 6:1 doi:10.1186/1472-6793-6-1 |
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