Mouse hind limb tibialis anterior (TA) muscles were injected with BaCl2 followed by daily systemic administration of rapamycin (Rap) starting 7 days after injury. The animals were sacrificed at different days, and the injured muscles were dissected and cryosectioned followed by hematoxylin (blue) and eosin (pink to red) staining. This image shows the regeneration of muscle tissue fourteen days after the injury with daily systemic administration of Rap from day 7 to day 13. The growth of muscle fibers, stained by eosin and showed as red patches, was impaired as indicated by the smaller average cross-section area of the fibers. Nuclei were stained by hematoxylin in blue. This image corresponds to Fig 10B from JCB 189: 1157-1169, 2010. See also CIL: 13595, 13596, 13597, 13599, and 13600.
TA muscles were isolated by dissection, frozen in liquid nitrogen–cooled 2-methylbutane, and embedded in TBS tissue freezing medium. Sections of 10-μm thickness were made with a cryostat at -20°C, placed on uncoated slides, and stained with hematoxylin and eosin. The stained slides were examined with a microscope (DMI 4000B), and the images were captured with a Fluotar 20× 0.4 NA dry objective (Leica) using a camera (RETIGA Exi). The images were processed as 24-bit colored images using Photoshop.
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