Volume 7 issue 7 July 2006
Dynamin is involved in endolysosomal cholesterol delivery to the endoplasmic reticulum: role in cholesterol homeostasis
Peggy Robinet, Alexandre Fradagrada, Marie-Noëlle Monier, Marta Marchetti, Anne Cogny, Nicole Moatti, Jean-Louis Paul, Benoit Vedie and Christophe Lamaze

Supplementary Figure 1: HeLa cells reach a steady-state for total cell cholesterol and sterol sensitive genes expression after 24 hours of growth in LPDS.
A) K44A HeLa cells were grown in LPDS medium for the indicated times and levels of LDL receptor, HMGCoA reductase, and SREBP-2 genes were expressed as fold variation over control at To after normalization. Each point represented the mean ± SEM of 3 independent experiments. All Ct determination were made in duplicate. B) K44A HeLa cells were grown in LPDS medium for the indicated times and total cholesterol was quantified as described in Materials and Methods. Each value is the mean ± SD of duplicate experiments and expressed as nanomoles per mg of cell proteins.

Figure 1 (.jpg)

Supplementary Figure 2: Expression levels of LDL receptor, HMGCoA reductase, and SREBP-2 genes in dynamin-inactivated cells after FC/25HC supplementation.
K44A HeLa cells expressing the K44A dynamin mutant (dynK44A) or not (control) were grown in LPDS medium for 24 h and further incubated with a combination of FC/25HC for 24 h. Expression levels of LDL receptor, HMGCoA reductase, and SREBP-2 genes were measured at 3 h and 24 h after the addition of FC/25HC. Results are expressed as fold variation over control at T0 after normalization. Each bar represents the mean  ± SD of triplicate experiments.

Figure 2 (.jpg)