Interaction between LXXLL peptides and PPARα in
mammalian two-hybrid assay. COS-1 cells were grown in serum-free
media for 12 hours before transfection with GAL4DBD(pM)/peptide
fusions, VP16/PPARα, pFR-luciferase, and pRL-TK. Panel (a):
cells were treated with 100 μM α-linolenic acid
(ALA), linoleic acid (LA), docosahexaenoic acid (DHA), conjugated
linoleic acid (CLA), octanoic acid (OCT), oleic acid (OA),
25 μM Wy-14,643 (Wy), 25 μM ciprofibrate
(Cipro), or DMSO (0.1% v/v) for 6 hours. Panel (b): cells
were transfected and treated as described above with DMSO, CLA,
Wy, and also with 100 μM eicosatetraynoic acid (ETYA),
bezafibrate (Beza), or clofibrate (Clo). Luciferase activity was
measured and corrected for extraction yield and transfection
efficiency (n = 3). Data was expressed relative to that of the
pM-empty construct and the presented heat map was generated in
GeneSpring (Agilent, Palo Alto, CA). Darker shading represents
higher luciferase activity.