Fig 1.
NR12S distributes between the PM and ERC.
(A) Receptor-mediated endocytic pathway in non-polarized mammalian cells. Transferrin and its receptor are shown as an example. EE, early endosome; SE, sorting endosome; ERC, endocytic recycling compartment; LE, late endosome. (B) Representative maximum projection images of NR12S labeling in U2OS-SRA cells. The ERC is labeled with Alexa405-Tf. Scale bar is 10 μm.
Fig 2.
Metabolic modulation of cellular cholesterol levels.
GC/MS measurement of free cholesterol levels in control, cholesterol depleted and overloaded U2OS-SRA cells. Cellular lipids were extracted and analyzed by GC/MS. Data represent averages (± SE) of three independent experiments normalized to control value. ** p < 0.01.
Fig 3.
(A) Representative maximum projection images of NR12S labeling in control, cholesterol depleted, and cholesterol overloaded U2OS cells. Scale bar is 10 μm. (B) Representative planes used in the analysis of NR12S fluorescence intensity in the PM, ERC and Cell contact regions. Areas of cell contact points used in analysis are boxed. Scale bar is 10 μm. (C) Analysis of NR12S green/red ratio of the ERC and plasma membrane. NR12S green/red ratio was normalized to control PM. Data represent averages (± SE) of four independent experiments of at least 15 cells per experiment; NS, non-significant; *** p < 0.001 compared to control PM.