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. 2022 Nov 28;13:1057413. doi: 10.3389/fphys.2022.1057413

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Copyright © 2022 Heymann, Paul, Huang, Kinold, Dietrich and Aberle.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Side alleles carrying missense mutations express full-length Side.(A-D) Confocal images of ShGFP control and side homozygous mutant embryos (stage 14, lateral views) stained against Side (magenta). (A) In controls, Side is strongly expressed in developing sensory neurons and their outgrowing axons (arrow). (B) Expression level and subcellular location of Side are similar in side ^I306 and controls. (C) In side ^H143 mutants, Side is restricted to the soma and axonal localization is abolished (arrowhead). (D) Side is not detected in side ^C137. Confocal images in this and the following figures are all maximum intensity projections. Anterior is left and dorsal is up. Scale bar: 25 μm. (E) Western blot of lysates from control and homozygous side mutant embryos developed with anti-Side antibodies. (Upper panel) Full-length Side migrates as a single band at 120 kDa. Lane 1: Exogenous, untagged Side is strongly expressed by Mef2-Gal4. Lanes 2–3: Embryo collections of all stages exhibit weaker Side signals than collections enriched for older embryos (st. 16–17, hand-sorted). Lanes 4–5: In side ^I306 and side ^H143, Side migrates at comparable size and expression level as endogenous Side. Lanes 6–9: Side is not detectable in side alleles carrying protein truncating mutations. Asterisk marks a suspicious band of unknown origin at approximately 47 kDa that cross-reacts with the monoclonal antibody and that seems to be enriched in all mutants. (Lower panel) The same blot developed with anti-α-Tubulin antibodies (loading control).