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Figure 13 | BMC Immunology

Figure 13

From: Discriminators of mouse bladder response to intravesical Bacillus Calmette-Guerin (BCG)

Figure 13

Chromatin immunoprecipitation (CHIP)/quantitative real-time polymerase chain reaction (Q-PCR)-Based Assays. Female C57BL/6J mice were anesthetized and instilled with 200 μl of one of the following substances: BCG (TheraCys®-Aventis-Pasteur; total dose of 1.35 mg) or pyrogen-free saline on days 1, 7, 14, and 21, as described in methods. Mice were euthanized 24 hours after a single instillation (BCG Acute) or 7 days after 4 weekly instillations (BCG Chronic). A total of 60 mice were used (20 mice per group). Bladders were removed rapidly, frozen, and shipped to Genpathway http://www.genpathway.com/ for querying the chromatin for transcription of listed genes (Genpathway's TranscriptionPath Query assay). After isolation, the chromatin was incubated with an antibody against RNA polymerase II (Abcam) to precipitate the DNA transcriptome. The final CHIP DNAs were then used as templates in Q-PCR reactions using specific primer pairs (Table 1). Results are presented as average and standard error of Transcription Events Detected Per 1000 Cells. Results were grouped to facilitate their visualization. IL-17 family. Graphics illustrated in figures 11, 12, and 13 present the same X-axis to permit comparisons between gene families.

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