The inflammatory and normal transcriptome of mouse bladder detrusor and mucosa

Saban, Marcia R; Hellmich, Helen L; Turner, Mary; Nguyen, Ngoc-Bich; Vadigepalli, Rajanikanth; Dyer, David W; Hurst, Robert E; Centola, Michael; Saban, Ricardo

doi:10.1186/1472-6793-6-1

BMC Physiology

Table 4 Differential expression of selected SSH transcripts by quantitative real-time polymerase chain reaction (QRT-PCR) ***

From: The inflammatory and normal transcriptome of mouse bladder detrusor and mucosa

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 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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ISSN: 1472-6793

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