Figure 10

NF-kappaB electrophoretic mobility shift assays (EMSA). Anesthetized C57BL6 female mice were 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 above. Mice were euthanized with pentobarbital (200 mg/kg, i.p.) 24 hours after a single instillation (BCG acute) or 7 days after 4 weekly instillations (BCG chronic). Bladder mucosa nuclear extracts were incubated with 2 ng of [³²P] NF-kappaB double-stranded probe. Supershift reactions were pre-incubated for 20 minutes with antibody against p65 or p50. Insert graph represents the quantification of p50/p65 NF-kappaB activity (indicated by red arrow) using ImageJ Software. Statistical differences were determined using GraphPad Software.