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. 2020 Feb 1;34(3-4):194–208. doi: 10.1101/gad.331330.119

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© 2020 Monahan Vargas et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 5. — Ringer and futsch operate in the same genetic pathway to promote axon regeneration, and Western blotting of ringer and Ac-Tub in various mutants. (A) Genetic interaction analysis between ringer and futsch. While C4da neuron axons in heterozygotes of futsch^N94/+ or ringer^915/+ behave similarly to WT, significant reduction of regeneration is observed in homozygotes of futsch^N94 and transheterozygotes of futsch^N94/+ and ringer^915/+ (futsch^N94/+; ringer^915/+). The injury site is demarcated by the dashed circle. Arrow marks axon stalling while arrowheads show the regrowing axon tips. (B,C) Quantifications of C4da neuron axon regeneration with regeneration percentage (B) and regeneration index (C). n = 20 to 64 neurons from five to 16 larvae. (D,E) Ringer and Ac-Tub levels in WT and mutants of ringer, futsch, and HDAC6. (D) Representative immunoblots showing total levels of ringer (left) and Ac-Tub (right) from larval lysates of specified genotypes. Actin was used as the loading control. (E) Quantification of the ratio of band intensities of ringer (left) and Ac-Tub (right) with respect to Actin in represented genotypes. n = 3 independent experiments. (*) P < 0.05; (**) P < 0.01; (***) P < 0.001 by Fisher's exact test (B), one-way ANOVA followed by Holm-Sidak's test (C), or Dunnett's test (E). Scale bar, 20 μm.