Images were acquired with B-mode, and color Doppler modes using a Vevo 2100 high frequency ultrasound machine (VisualSonics) and a MS550D 22–55 MHz probe. Animals were sedated with isoflurane and scan time was approximately 20 min to identify and image the spleen. Spleen area was calculated from 2-dimensional images using the VisualSonics software.
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