Fig. 6

Fluorescent fatty acid tracers reveal NDRG1 suppresses fatty acid incorporation Into neutral lipids and storage in lipid droplets. a Fluorescence micrographs of stable NDRG1 SKBR3 +/− NDRG1 depletion fed the 16-carbon boron-dipyrromethene (BODIPY) palmitate (BODIPY C16–100 μM, 16 h), fixed. Lipid droplets and nuclei counterstained with LipidTox Deep Red and Hoechst, respectively. b Quantification of total BODIPY C16 uptake in SKBR3 cells +/− NDRG1 depletion. Graphs represent average total signal per cell at indicated tracer concentrations. c Quantification of lipid droplet specific BODIPY C16 by segmentation of small granules. d Relative distribution of BODIPY C16 signal representing droplet/whole cell signal. e LipidTox stained lipid droplet numbers represented as objects per cell. f Comparison of lipid droplet intensities of starved and fed controls at day 3 in stable BODIPY 558/568 C12 loaded MCF7 cells. Complete medium versus Hank’s buffered saline (HBSS). g BODIPY 493/503 counterstaining of NDRG1-FLAG overexpressing and control cells fed complete medium or HBSS for 3 days. h Analysis of cell survival due to starvation in stable BODIPY 558/568 C12-fed MCF7 cells transduced with NDRG1-FLAG or empty vector after 3 days of culture in HBSS. i, j Time course of changes in lipid droplet formation and cell count due to HBSS starvation in stable MCF7 cells transduced with the indicated constructs. Values represent HBSS/complete medium. All bar graphs represent mean +/− SD and comparisons were by two-sided t test. N = 12/group for all. Scale bars = 25 μm. CTRL, control