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The final stage is the disappearance of the tail, but this takes place rather later, the tissue being used to produce a spurt of growth in the limbs.
Five days later, the tissue blocks were removed, and the medium was replaced.
Later, the tissue blocks were cut into 5-μm sections, placed onto glass slides and stained with hematoxylin and eosin (H&E), dehydrated, and mounted.
Later, the tissue is rather characterized by supracellular F-actin fibers oriented roughly parallel to the closing embryonic flanks (Fig. 3I).
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Later, the tissues were permeabilized with 0.5% Triton X-100 for 10 minutes and blocked by 10% fetal bovine serum for 1 hour.
Twenty-four hours later, the tissues were embedded in resin.
Later the tissues were homogenized in 1.15 % KCl solution and 20%% (W/V) homogenate, which was later, centrifuged and then used to measure SOD activity using spectrophotometric method.
Twelve weeks later, the tissues including liver, spleen, heart, and aorta of the mice were taken out and rinsed with precooling normal saline.
RNA was isolated from the brain samples from mice stored in RNA later by spinning down and removing the RNA later from the tissue and then isolating RNA using the RNA-Bee protocol described above.
Animals were perfused 2 h later and the tissue processed as detailed above.
Biopsies were taken at 4.5 months of age and repeated two weeks later if the tissue was inadequate for a diagnosis.
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