For population measurements of nerve ring axon defects, animals were mounted on slides and visualized in an Axioscope compound microscope (Zeiss). Mutant phenotypes were quantified using axon markers to define nerve ring entry and growth based on the meeting point of the dorsal axon part of bilaterally symmetrical neurons. When scoring axons of ASE or AIY neurons, wild type animals show a full axonal ring with a dorsal meeting point of bilaterally symmetrical axons, while mutant animals present an axonal ring with a gap and no dorsal meeting point. Images presented in Fig. 3a–h are of L3–L4 animals for better visualization of the defect. Scoring of mutant phenotypes was performed in L in animals of early larval stages L1–L3; no significant difference of defects was observed in between those early larval stages.
For embryonic axon length measurements, images were acquired on a Deltavision microscope, as described below. Axons were traced and axon length was measured manually using image stacks in ImageJ/Fiji. Axon length was measured from the anterior limit of cell body fluorescence to the anterior limit of axon fluorescent tip.
For embryonic axon length measurements, images were acquired on a Deltavision microscope, as described below. Axons were traced and axon length was measured manually using image stacks in ImageJ/Fiji. Axon length was measured from the anterior limit of cell body fluorescence to the anterior limit of axon fluorescent tip.