Fig. 6.

Lack of correlation between MTP expression and frequency of nLDs and LAPS in intestinal-derived cells. A: cells were treated with oleate (500 μM) for 24 h and subsequently immunostained with PML and CCTα primary and AlexaFluor-647 and AlexaFluor-555 secondary antibodies, respectively. LDs were visualized with BODIPY 493/503 and imaged by confocal microscopy (bar, 10 μm). B: percentage of nLD- and LAPS-positive cells from a representative experiment (mean and SD). C: cell lysates were immunoblotted for MTP, CCTα, and actin (load control). D: average nLD cross-sectional area in Caco2 cells treated for 24 h with oleate or oleate plus MTPi. Results are from a representative experiment using 8–10 fields of cells (30–60 cells/field). E: xyz-projection of mouse enteroids immunostained for CCTα and LMNA/C. LDs were visualized with BODIPY 493/503 and nuclei stained with Hoechst. The panels to the right are individual channels from the xy-plane showing the CCTα-positive nLD (indicted by arrow) in the nucleus of an enterocyte (bar, 5 μm). CCTα, CTP:phosphocholine cytidylyltransferase; LAPS, lipid-associated promyelocytic leukemia structure; MTPi, MTP inhibitor; nLD, nuclear lipid droplet; PML, promyelocytic leukemia.