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. 2021 Jan 21;10:e62232. doi: 10.7554/eLife.62232

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© 2021, Wagstaff et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 2. — (A) Analysis of RNA sequencing data showing decrease in gene expression during chronic denervation. (B) Representative western blot showing p75NTR expression in uninjured (UI) nerves and distal nerve stumps following 3 days, and 1, 3, 6, 8 and 10 weeks of denervation. The graph shows quantitation of the results. P75NTR peaks 1 week after injury and gradually declines during prolonged denervation. Data normalized to 1 week after injury. One-way ANOVA with Dunnett's multiple comparison test; **p<0.005, ***p<0.001. n = 4. (C) Counts of back-filled Fluorogold-labeled regenerating motor neurons following immediate repair or chronic 10-week denervation show a decrease in motor neuron regeneration into chronically denervated stumps. Unpaired Student’s t-test; **p=0.0020. n = 6 for each time point. (D) Counts of neurofilament+ axons mirrors the decline in regeneration observed with chronic denervation shown in C. Counts were performed on transverse sections taken 3 mm from the repair site1 week after repair. Unpaired Student’s t-test; ****p<0.0001. Immediate repair n = 5, chronic denervation n = 4. (E) Counts of back-filled Fluorogold-labeled motor neurons showing similar numbers of regenerating neurons following immediate repair or repair after 1 week of denervation. Unpaired Student’s t-test; p=0.9. n = 3. All numerical data represented as means ± SEM.

Figure 2—figure supplement 1. — (A) Analysis of RNA sequencing data showing decrease in gene expression during chronic denervation. (B) Representative western blot showing p75NTR expression in uninjured (UI) nerves and distal nerve stumps following 3 days, and 1, 3, 6, 8 and 10 weeks of denervation. The graph shows quantitation of the results. P75NTR peaks 1 week after injury and gradually declines during prolonged denervation. Data normalized to 1 week after injury. One-way ANOVA with Dunnett's multiple comparison test; **p<0.005, ***p<0.001. n = 4. (C) Counts of back-filled Fluorogold-labeled regenerating motor neurons following immediate repair or chronic 10-week denervation show a decrease in motor neuron regeneration into chronically denervated stumps. Unpaired Student’s t-test; **p=0.0020. n = 6 for each time point. (D) Counts of neurofilament+ axons mirrors the decline in regeneration observed with chronic denervation shown in C. Counts were performed on transverse sections taken 3 mm from the repair site1 week after repair. Unpaired Student’s t-test; ****p<0.0001. Immediate repair n = 5, chronic denervation n = 4. (E) Counts of back-filled Fluorogold-labeled motor neurons showing similar numbers of regenerating neurons following immediate repair or repair after 1 week of denervation. Unpaired Student’s t-test; p=0.9. n = 3. All numerical data represented as means ± SEM.