Journal: The Journal of Biological Chemistry
Article Title: A Block in Endoplasmic Reticulum-to-Golgi Trafficking Inhibits Phospholipid Synthesis and Induces Neutral Lipid Accumulation *
Figure Lengend Snippet: Fluorescence microscopic analysis of lipid droplets. Prior to microscopy, cultures were grown in I + medium at 25 °C as described under “Experimental Procedures” and were then diluted to A 600 nm = 0.1 in 2 ml of I + medium in multiwell plates, and incubated at 25 °C in an Eppendorf incubator/mixer. After reaching A 600 nm = 0.5, cultures were shifted to 37 °C, and aliquots removed at time 0 and 120 min, fixed and stained with BODIPY® 493/503, and analyzed by fluorescence microscopy. a , wild type cells at 25 °C (zero time at 37 °C); b , after 120 min of incubation at 37 °C; c, sec13-1 cells at 25 °C (zero time at 37 °C); d , after 120 min of incubation at 37 °C; e, sec13-1lro1 Δ dga1 Δ are 2Δ cells at 25 °C (zero time at 37 °C); and f , after 120 min incubation at 37 °C. Note that fluorescence emitted from BODIPY® that is dissolved in membrane aggregates, as opposed to lipid droplets, is r ed shifted, as indicated by a more yellowish fluorescence in e and f. Insets are 2-fold magnifications of representative cells. Bar = 5 μm.
Article Snippet: They were then diluted once more to A 600 nm = 0.1 in 2 ml of I+ media in multiwell plates and incubated at 25 °C in an Eppendorf incubator/mixer.
Techniques: Fluorescence, Microscopy, Incubation, Staining