Nem1p localizes next to lipid droplets on the ER. (A) Nem1p-mCherry was chromosomally expressed at the NEM1 locus. Lipid droplets were stained with BODIPY. (top left) Brightfield image. (bottom left) Projection image of BODIPY-stained cells showing Nem1p-mCherry localization (bottom left inset). The arrowhead is an example of colocalization of CFP-HDEL (ER), BODIPY (lipid droplet), and Nem1p-mCherry. Bars, 5 µm. (right) Three-dimensional reconstruction of the same field of cells. Green, droplets; pink (10 dots), Nem1p within 0.65 µm of a droplet; blue (two dots with arrowheads), Nem1p-mCherry beyond 0.65 µm of a droplet. (B) Juxtaposition of Nem1p-mCherry puncta with lipid droplets (LD). Nem1p-mCherry puncta were counted from seven separate groups of cells and scored for their observed association with lipid droplets. Mean ± SEM was calculated across all seven groups.