Sodium azide

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).

Hemerocallis citrina Borani was obtained from Qingyang, Gansu, China. Standards of glucose, mannose, fucose, rhamnose, xylose, galactose, arabinose, glucuronic acid, ribose, and galacturonic acid were purchased from Shanghai Yuanye Biotechnology (Shanghai, China). Absolute ethanol was purchased from Fuyu Fine Chemical (Tianjin, China). Trifluoroacetic acid, methanol, sodium hydroxide, hydrochloric acid, carbazole, 1-phenyl-3-methyl-5-pyrazolone, 3,5-dinitrosalicylic acid, sulfuric acid, Coomash bright blue, chloroform, potassium bromide, sodium azide, sodium acetate, anthranone, and other reagents were purchased from Solarbio Biotechnology (Beijing, China). ABTS and FRAP assay kits were purchased from Suzhou Keming Biotechnology (Suzhou, China). Distilled water was used in these experiments.