Technical Reference #414

414.

Regulated Step In Cholesterol Feedback Localized To Budding Of Scap From Er Membranes Axel Nohturfft; Daisuke Yabe; Joseph L. Goldstein; Michael S. Brown; and Peter J. Espenshade, University of Texas, Cell, 102(414), (2000)

Abstract:
SREBPs exit the ER in a complex with SCAP. Together they move to the Golgi where SREBP is cleaved re-leasing a fragment that activates genes encoding lipid biosynthetic enzymes. Sterols block ER exit pre-ventingcleavage decreasing transcription and

Keywords:
SCAP; ER Membranes; Sterols; Cholesterol homeostasis; Sterol Regulatory Element Binding Proteins (SREBPs); cleavage

Materials & Methods:
On day 0 cells were set up in glass-bottomed 35 mm dishes (MatTek Co. Ashland MA) at a density of 3 3 104 cells/dish in medium A supplemented with 5% FCS. On day 2 cells were washed with PBS and refed 1 ml of medium A supplemented with 5% newborn calf

Microscopic Technique
Confocal Microscopy, Time Lapse

Cell Type(s)
CHO-K1