Segregation of HPos into LD after lipid arrival. (A) Starved GFP-HPos–transfected cells were treated with OA for 7.5 min and analyzed by microscopy. (B) Starved OFP-HPos– and GFP-HNeu–cotransfected cells were loaded with OA for 7.5 min and analyzed by microscopy. The inset shows a high-magnification detail of an emerging LD. (C and D) Starved GFP-HPos–transfected cells were loaded with OA for 30 min. The distribution of neutral lipids was detected with Nile red (red in C), and the distribution of endogenous Plin3 was detected with antibodies (red in D). Asterisks indicate transfected cells. Bars: (A, C, and D) 20 µm; (B, main panel) 2 µm; (B, inset) 0.5 µm.