Fig 6.

Elovl1 synthesizes C₂₆-CoA in cooperation with CerS3 in the SG. (A) Skin samples isolated from wild-type mice at E18.5 were fixed with 4% paraformaldehyde, hybridized with a digoxigenin-labeled CerS2 or CerS3 RNA probe, and stained with alkaline phosphatase-conjugated antidigoxigenin antibody (F_ab fragment) and nitroblue tetrazolium–5-bromo-4-chloro-3-indolyl phosphate solution. Frozen sections (20 or 25 μm) were subjected to microscopic observation under a DM5000B light microscope and photographed. Bar, 50 μm. De, dermis. (B) HEK 293T cells transfected with control vectors or plasmids carrying ELOVL1 and CERS2 or CERS3 were labeled with 2 μCi of [³H]sphingosine for 2 h at 37°C. Lipids were extracted, separated by reverse-phase TLC, and detected by autoradiography. Positions of Cer species are indicated on the left.