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. Author manuscript; available in PMC: 2016 Mar 23.
Published in final edited form as: Dev Cell. 2015 Mar 5;32(6):678–692. doi: 10.1016/j.devcel.2015.01.029

Figure 1. Fatty acid trafficking can be visualized using a fluorescent fatty acid pulse-chase assay.

Figure 1

(A) Schematic representation of the fluorescent FA pulse-chase assay: cells were pulsed with Red C12 overnight, washed, and incubated with CM for 1 h in order to allow the Red C12 to accumulate in LDs. Cells were then chased in CM or HBSS for the indicated periods of time and imaged to determine the subcellular localization of the FA. (B–D) WT MEFs were assayed as described in Figure 1A and chased in CM or HBSS for 0 h, 6 h, or 24 h. LDs were labeled using (B) BODIPY 493/503 and (C) mitochondria were labeled using MitoTracker Green. Scale bar, 10μm. (D) Relative cellular localization of Red C12 was quantified by Pearson’s coefficient analysis. Data were expressed as means ± SEM. (E) TLC resolving Red C12 isolated from WT MEFs assayed as described above and chased for 6 h or 24 h with HBSS in the absence or presence of etomoxir, see also Figure S1.