Spatial Distribution of Hc-TTR-31 in C. elegans

To confirm the spatial distribution of Hc-TTR-31, transgenesis of Hc-ttr-31 was conducted in C. elegans (Huang et al., 2021 (link)). Briefly, the promoter sequence of Ce-ttr-31 was amplified, ligated to the PMD-18T vector and subcloned into pPD95.67 plasmids (Ce-ttr-31p::gfp), with the Hc-ttr-31 coding sequence inserted into the multiple cloning site (Ce-ttr-31p::Hc-ttr-31::gfp). Transgenic expression of Hc-ttr-31 in C. elegans N2 strain was performed by micro-injection with the Ce-ttr-31p::Hc-ttr-31::gfp (Mello et al., 1991 (link)). A Ce-ttr-31p::Ce-ttr-31::gfp was used as a reference control. Considering the homology of Hc-ttr-31 and Ce-ttr-31, transgenesis of Hc-ttr-31 should result in an over-expression of TTR-31 in the treated C. elegans. To explore the effects of exogenous Hc-TTR-31 in C. elegans, transgenic worms were mounted on 2% agar pads containing 1% sodium azide (Solarbio, China) and imaged using a Zeiss LSM710 laser confocal microscope (Zeiss Microscopy).