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. 2019 Jun 21;218(8):2583–2599. doi: 10.1083/jcb.201902061

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© 2019 Chang et al.

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Figure 6. — Monitoring LD-to-peroxisome FA trafficking using NBD-C12. (A) Schematic diagram representing NBD-C12 pulse-chase assay using with DEUP treatment. (B) Distribution of NBD-C12 in BODIPY-665/676–labeled LDs and mCherry-SKL–labeled peroxisomes after pulse (top) and after pulse-chase in control (middle) or 150 µM DEUP-treated HeLa cells (bottom). Magenta and red arrowheads indicate LDs and peroxisomes, respectively. Representative confocal MIP images are shown. (C) Relative NBC-C12 intensity in LDs or peroxisomes as described in B. Means ± SEM are shown (26–27 cells from three independent experiments). **, P < 0.01; ***, P < 0.001. (D) TLC analysis of NBD-C12 extracts from pulse-chased HeLa cells as described in A. C12, 1 µl of 1 µM NBD-C12 loading for identifying free form of C12 from cell extracts. (E) TLC analysis of NBD-C12 extracts from siCtrl and siABCD1 transfected HeLa cells.