Screening Natural Compounds for SREBP2 Activation

Luciferase assay was performed as previously described [1 (link)]. HEK293T cells transiently transfected with steroid responsive element (SRE)-wild type (WT) or -mutant (Mut) containing luciferase vector (pSynSRE-T-Luc and pSynSRE-Mut-T-Luc) by using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). pSynSRE-T-Luciferase vectors (Addgene plasmid #60444 and #60490) were gifts from Timothy Osborne [66 ]. To screen the SREBP2 activating natural compounds, transfected HEK293T cells were incubated with a library of 502 natural compounds (20 μM of each compounds) for 24 h. Luciferase activities were analyzed by using a Synergy 2 Luminometer (BioTek, Winooski, VT, USA) and β-gal assay was used for normalization.