Figure 11

SFaN does not interact with SREBP-1a. (A) Structure of alkynyl-SFaN. (B) Huh-7/FAS cells were sterol-depleted by incubation in medium C for 16 h. The cells were then switched to medium C in the presence of the vehicle or the indicated concentrations of SFaN or alkynyl-SFaN. After incubation for 24 h, a luciferase assay was performed, and relative luciferase activity was obtained by activity normalization in the presence of the vehicle. All data are represented as the means ± S.E. (n = 3). Different superscript letters denote statistical significance (p < 0.05). (C) Huh-7 cells were sterol-depleted by incubation in medium D for 16 h. The cells were then shifted into medium D in the presence of the vehicle or the indicated concentrations of alkynyl-SFaN. After incubation for 3 h, whole-cell extracts underwent immunoblotting (IB) with anti-SREBP-1 (2A4), anti-SREBP-2, or anti-β-actin antibodies. (D) Structure of SFaN beads. (E) HEK293 cells were transfected with pCMV-SREBP-1a-(2–1147)-3 × FLAG and pCMV-Keap1-3 × FLAG and cultured in medium B for 24 h. Whole-cell extracts underwent pull-down experiments with SFaN beads with or without the addition of 100 μM SFaN and were then subjected to immunoblotting (IB) using an anti-FLAG antibody. Full-length images are presented in Supplementary Fig. S10.