Transurethral instillation was performed as previously described [7] (link). 8 week old C3H/HeOuJ male mice (Jackson Laboratories) were anesthetized with isoflurane and catheterized with a lubricated sterile polyethylene catheter per urethra. Inoculation was performed by a single transurethral instillation of uropathogenic E. coli 1677 (2×106 CFU/ml) or sterile PBS in a volume of 200 µl.
Animals were sacrificed 2, 7, 14, 21, and 28 days post-instillation. The paired prostatic lobes (VP, DLP, AP) were bisected separately and were used for RNA isolation, hydroxyproline assay, and/or histology. The whole mouse prostates were collected and used for measuring incorporation of 3H-hydroxyproline to determine collagen synthesis. Four to thirteen mice per time point and per treatment were analyzed for histology. Four to seven mice per time point and per treatment were used for gene expression analysis. Four to eight mice per time point and per treatment were used for hydroxyproline assay. Four to seven mice per time point and per treatment were used for determination of collagen synthesis. Four to six mice per treatment were used for FACS analysis.
Animals were sacrificed 2, 7, 14, 21, and 28 days post-instillation. The paired prostatic lobes (VP, DLP, AP) were bisected separately and were used for RNA isolation, hydroxyproline assay, and/or histology. The whole mouse prostates were collected and used for measuring incorporation of 3H-hydroxyproline to determine collagen synthesis. Four to thirteen mice per time point and per treatment were analyzed for histology. Four to seven mice per time point and per treatment were used for gene expression analysis. Four to eight mice per time point and per treatment were used for hydroxyproline assay. Four to seven mice per time point and per treatment were used for determination of collagen synthesis. Four to six mice per treatment were used for FACS analysis.