ORP5 localizes to nascent LDs and the preexisting ones in Huh7. (A) Airyscan video snapshots of Huh7 cells expressing EGFP-ORP5B (green), RFP-Sec22b (red) and Mito-BFP (blue). Cells were treated with OA to induce the formation of LDs, and stained by LTox (purple). Images were taken every 5 min for 50 min. Scale bar, 10 μm (entire cell), or 5 μm (zoom). (B) Another example depicting the recruitment of ORP5 to pre-existing LDs, with mitochondria. Arrowheads indicate EGFP-tagged ORP5 at MAM-LD contacts. Scale bar, 10 μm (entire cell), or 2 μm (zoom). (C) Line graph shows the distribution of the nearest-neighbor EGFP-ORP5B puncta to LDs in the images of Huh7 cells treated with OA. The observed probability density largely deviates from randomly distributed LD and puncta EGFP-ORP5B. (D) Airyscan video snapshots of Huh7 cells expressing EGFP-ORP5B (green), Sec61β-mCherry (red) and Mito-BFP (blue). Cells were treated with OA to induce the formation of LDs, and stained by LTox (purple). Scale bar, 10 μm (entire cell), or 5 μm (zoom).