Fig. 6: SREBP-1c cleavage-activation is suppressed by lipolysis derived uFAs.

A Primary hepatocytes from wild type mice were transfected with pGFP-SCAP one day after isolation. After 48 h, cells were treated with 1% w/v HPCD [(2-hydroxypropyl)-beta-cyclodextrin] for 1 h. Thereafter, all cells were incubated in 5% LPDS medium with mevalonate and mevastatin present (lipid depletion), with the addition of 100 μM 16:0 or 18:1 FA or with the addition of 2.5 μM of 25-hydroxycholesterol (25-HC), as indicated. 2 h later, cells were fixed, permeabilized, and GFP-SCAP was visualized by anti-GFP immunofluorescence (IF) (green); Golgi was imaged by anti-GM130 IF (red); DAPI was used for nuclear staining. B SCAP-GFP/GM-130 colocalization was determined by Pearson’s correlation coefficient, see dot-plots. Unpaired t-tests were used to compute significance levels, n = 40–46 cells/group.