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Fig 1.

Schematic view of amputation site and parameters analyzed.

(A) Representative amputation area of the rat limb and stump samples collected for analysis. (B) Schematic description of parameters analyzed.

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Fig 2.

Specific NF staining of nerve regrowth and neuroma development observed in the tissue sections from day 3 until 90.

(A, B, C, D and E) Schematic figures representing an overview of structures location and representative images of axonal fascicles show larger and closer view (a, b, c, d and e with 4x magnification; ai, bi, ci, di and ei with 20x magnification, scale bar = 100μm) of structures throughout time.

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Fig 3.

Axonal density of normal and degenerating axons observed in all time points.

Boxplot overlaid with sample distribution of axonal density of normal and degenerating axons. * Statistically significant differences (p≤0.05).

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Fig 4.

Classification of histological characteristics of nerves in limb stumps after amputation.

Normal nerve (N0i: longitudinal and N0ii: transversal section), degenerating axons (N1), axonal sprouts (N2i: at the end of damaged fascicles, arrows show elongated structures corresponding to axonal fibers and N2ii: at distal area, arrows show isolated axons appearing as donut ring-like structures), unorganized axons bundled in connective tissue (N3), unorganized axon ramifications in muscle (N4), neuroma—highly unorganized axons in tissue (N5). Scale bar = 100μm.

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Fig 5.

Prevalence of morphological characteristics in rat limb stump tissue analyzed at each time point.

Error bars represent confidence interval of 95%.

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