Fig. 2.

CCTα knockout in undifferentiated Caco2 cells results in fewer and larger nLDs. A: representative confocal images of Caco2 and CCTα-KO cells treated with oleate (400 μM) for 24 h used for quantitation in panels B–G (12 h images not shown). Cells were immunostained with PML and LMNA/C primary followed by AlexaFluor-594 and AlexaFluor-647 secondary antibodies, respectively (bar, 5 μm). LDs were visualized with BODIPY 493/503. B: quantification of cLDs per cell. C: quantification of nLDs per cell. D: the mean area of nLDs. E: binned distribution of nLD area in oleate-treated cells. F: ratio of nLDs to total cLDs plus nLDs. G: percentage of LAPS (PML+ve nLDs) in oleate-treated cells. Results are presented as scatter plots showing the mean and SD from 6-12 fields of cells (10–15 cells/field) from three separate experiments. Panel E is box and whisker plots showing the mean and 5th-to-95th percentile. Significance was determined by two-way ANOVA. ∗∗∗P < 0.001, ∗∗P < 0.01. CCTα, CTP:phosphocholine cytidylyltransferase; cLD, cytoplasmic lipid droplet; LAPS, lipid-associated promyelocytic leukemia structure; nLD, nuclear lipid droplet; PML, promyelocytic leukemia.