E1003 triglyceride assay kit

African green monkey kidney fibroblast-like COS-7 cells were purchased from the Cell Center of the Chinese Academy of Medical Sciences (China). Human neuroblastoma SK-N-SH and SK/NEST cells that stably expressed NEST were from our previous study [44 (link)]. The NTE-GFP and ΔR-NTE-GFP constructs were generous gifts from Dr. Paul Glynn [6 (link)]. Plasmid pEGFP-N3 was purchased from Clontech (Mountain View, CA, USA). Transfection reagent Lipofectamine 2000 and HCS LipidTOX™ Deep Red neutral lipid stain were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Cell culture reagents and OA were from Sigma-Aldrich (St. Louis, MO, USA). The E1003 triglyceride assay kit was purchased from Applygen Technologies (Beijing, China). The Q5 Site-Directed Mutagenesis Kit was purchased from New England Biolabs (Ipswich, MA, USA). Mouse anti-GFP, anti-GAPDH monoclonal antibodies, and goat anti-mouse IgG HRP were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Enhanced chemiluminescence (ECL) reagents were obtained from Beyotime Biotechnology (Shanghai, China).

Samples of unicellular organisms such as Saccharomyces cerevisiae, Escherichia coli, Pseudomonas syringae, Chlamydomonas reinhardtii and conidia of Trichoderma viride were cultured in appropriate culture conditions. Resting spores of P. brassicae were extracted from galls of rapeseed and then surface disinfested. The sample suspensions were harvested by centrifugation at 4,000 g for 10 min in a 50-ml tube, washed twice with 10 ml PBS, and lysed by compressing three times with a French press at 1,500 bar at 4 °C. Whole sample lysates were then centrifuged at 12,000 g for 5 min at 4 °C. The lysates were examined under a microscope to ensure that the cells were completely broken. The TAG content of the supernatant was measured using an E1003 triglyceride assay kit (Applygen Technologies, China), and the protein concentration was quantified using a Pierce BCA Protein Assay Kit (Thermo, USA).