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. Author manuscript; available in PMC: 2012 Aug 3.
Published in final edited form as: Cell Metab. 2011 Aug 3;14(2):173–183. doi: 10.1016/j.cmet.2011.06.008

Figure 4. Autophagy mobilizes hypothalamic lipids to generate endogenous fatty acids.

Figure 4

(A) BODIPY staining in GT1–7 cells cultured in presence (+S) or absence of serum (−S) for 30min or refed (RF) for 15min following serum removal. Values represents lipid droplet (LD) number per cell, and are mean+SE, for 20 different cells in each experiment, n=3. (B) Immunofluorescence for BODIPY/LAMP1 in GT1–7 cells in +S or −S for 30min or RF for 15min, and treated with or without lysosomal inhibitors (In) for 30min. Values represent % colocalization between BODIPY and LAMP1 (orange arrows), and are mean+SE for 20 different cells in each experiment, n=3. (C) Confocal images for BODIPY/LAMP1 in primary hypothalamic neurons co-treated with or without 0.25mM oleic (OL) or lysosomal inhibitors (In) for 4h. Values represent number of colocalized (orange arrows) puncta, and are mean+SE of more than 20 different cells in each experiment, n=3. See also Figure S4. (D) Thin layer chromatogram (TLC) of 14C-triglycerides (14C-TG) and 14C-fatty acids (14C-FA) from GT1–7 cells maintained in +S or −S and treated with or without triacsin C (TrC) for 30min. (E) TLC of 14C-TG and 14C-FA from TrC-treated GT1–7 cells in +S or −S for 30min or RF for 15min. Relative densitometric values represent 14C-FA per µg protein, and are mean+SE, n=3. (F) Scheme for experiment in E. (G) β-Oxidation assay in GT1–7 cells in +S or −S for 1h or RF for 30min. Values represent 14CO2 release expressed as counts per minute (CPM)/h/µg protein, and are mean+SE, n=3. *p<0.05, ***p<0.001.