RAG−/− mice were provided with autoclaved water supplemented with antibiotics [ampicillin (1 g/liter), metronidazole (1 g/liter), neomycin (1 g/liter), and vancomycin (0.5 g/liter)] for 6 days and then provided with autoclaved water for 1 day. Then, naive CD4+ T cells (CD4+CD25-CD62LhiCD44lo) from WT and RORγtcreCxxc1fl/fl mice were sorted and intravenously transferred into Rag1−/− mice at 2 × 106 cells per mouse. Two days later, the recipient mice were subjected to C. rodentium infection as described (39). Briefly, mice were gavaged with 5 × 108 C. rodentium cells in 250 μl of PBS per mouse. Bacteria were prepared by shaking at 37°C overnight in LB broth, and then, the cultures were serially diluted and plated to measure the colony-forming units. Body weight was measured daily. Fecal pellets were collected, weighed, and then homogenized in sterile PBS, and C. rodentium colonies were identified on the basis of morphology after 18 to 24 hours of incubation at 37°C on MacConkey agar plates.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.