Cba j females

The Institutional Animal Care and Use Committee at UCSF approved all procedures. The UCSF Laboratory Animal Resource Center (LARC) provided veterinary services for the study and all studies were performed in accordance with UCSF guidelines regarding animal housing, pain management, and euthanasia. All mice used were CBA/J females (Jackson Laboratory) aged between 8–10 weeks. LB cultures were inoculated with single colonies of E. coli and were placed in an oscillating incubator (~120 rpm) at 37 °C until they reached OD₆₀₀ of 1.0 (approximately 12–14h). For heat-killed samples, 1 mL aliquots of the culture were incubated at 90 °C for 30 minutes and then pelleted by centrifugation and resuspended in 100 μL of LB broth. Mice were placed under isoflurane anesthesia on a warming pad and 100 μL of live bacterial culture and 100 μL of 10X concentrated heat-killed bacteria were injected with tuberculin syringes into the right and left shoulder musculature, respectively. Mice were then removed from anesthesia, allowed to recover, and intermittently monitored prior to imaging. Imaging of E. coli-infected mice was initiated at approximately 8 hours following inoculation for [¹¹C]PABA studies.