Extended Data Fig. 9. Organelle distribution in knockout cells.
a, Simultaneous live imaging of six organelles with spectral unmixing. Two representative cells are shown. Note that where there is more ER, there tends to be more of the other organelles. b, Representative images of organelle distributions in CLIMP63 or p180 knockout cells. Markers used: mEmerald-Sec61β for ER; anti-TOM20 for mitochondria; anti-GM130 for Golgi apparatus; anti-EEA1 for endosomes; anti-LC3 for autophagosomes; anti-catalase for peroxisomes, anti-Lamp1 for lysosomes; and LD540 for lipid droplets. Cells labeled for autophagosome distribution were starved in EBSS for 2 h. ER is shown in green, while other organelles are in red. c, d, Quantifications of the distributions of different organelles labeled with specific markers in wild-type or else CLIMP63, KTN1, or p180 knockout cells. n = 30, 31, 30, 46 cells (left to right) for mitochondria MDR, n = 83, 86, 87, 46 cells for lipid droplet MDR, n = 30, 31, 31, 46 cells for peroxisome MDR, n = 29 cells for Golgi MDR, n = 30 cells for Autophagosome and lysosome MDR; n = 51, 46 cells for mitochondria asymmetry, n = 29, 31 cells for Golgi asymmetry, n = 49, 50 cells for lipid droplet asymmetry, n = 31 cells for autophagosome, lysosome and peroxisome asymmetry. e–g, Quantifications of lysosome, mitochondria, and peroxisome distributions in wild-type or p180/KTN1 double knockout cells transfected with control vector or CCP1. n = 32, 36, 36, 37 cells (left to right) for (e) and (f), n = 41, 48, 44, 40 cells for (g). Scale bars, 10 μm. All bars represent mean ± s.d. with individual data points shown. P values are shown along the top, two-tailed t-tests.