Imaging for luciferase expression in vivo was performed as previously described [29 (link)]. Briefly, animals were anesthetized with isoflurane (Abbott Laboratories), injected intraperitoneally (I.P.) with firefly D-luciferin (15 mg/mL in PBS; Gold Biotechnology, St. Louis, MO, USA), and imaged 5 min later with a cooled charge-coupled device (CCD) camera (IVIS; PerkinElmer, Waltham, MA, USA). The luciferin dose was 150 mg/kg; the volume varied with the age/size of the animal. Grey-scale photographs were acquired with a 24 cm field of view and a bioluminescence image was obtained using a binning/resolution factor of 4, a 1.2/f stop, and open filter. Regions of interest (ROIs) were defined manually using a standard area for each organ under investigation. Signal intensities were calculated with Living Image software v4.0(Perkin Elmer) and expressed as photons/second/cm2/steradian.
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