Skip to main content
. 2023 Oct 11;64(11):100457. doi: 10.1016/j.jlr.2023.100457

Fig. 6.

Fig. 6

Subcellular localization of cytosolic lipases in Neuro-2a cells and primary cortical neurons. Cells were incubated with oleic acid to induce lipid droplet (LD) formation. A: Neuro-2a perinuclear supernatants (PNS) were separated into cytosol (Cyt), membrane (Mem), and LD fractions by ultracentrifugation, and protein distribution was assessed by Western blotting analysis. B: subcellular localization of recombinant lipases in living Neuro-2a cells. Cells transiently expressing ATGL-ECFP or DDHD2-ECFP were incubated with oleic acid to induce LD formation and imaged by confocal fluorescence microscopy. LDs were detected using LipidTOX. Scale bars full size: 10 μm; scale bars insets: 1 μm. C, D: subcellular localization of endogenous DDHD2 in (C) Neuro-2a cells and (D) primary mouse neurons. Cells were incubated in the presence of oleic acid and in the absence or presence of KLH45 as indicated. Cells were fixed, and DDHD2 was detected by immunocytochemistry. LDs and cis-Golgi were detected using LipidTOX and anti-GM130 antibody, respectively. Scale bars full size: 10 μm; scale bars insets: 1 μm. ATGL, adipose triglyceride lipase; DDHD2, DDHD domain-containing 2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Ire1α; inositol-requiring enzyme 1α; PLIN2, perilipin 2.