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. 1998 Jun 29;141(7):1489–1502. doi: 10.1083/jcb.141.7.1489

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Localization and dynamics of ER/Golgi SNAREs by subcellular fractionation. NRK cells were incubated at 37°C or 15°C for 2 h, and were then homogenized and fractionated on sucrose equilibrium density gradients. (A, D, G) 0.75–1.75 M sucrose gradients that resolve Golgi/IC membranes (fractions 2, 3, and 4) from ER membranes (fractions 8, 9, and 10). Material that pelleted during centrifugation is included with fraction 14. (B, C, E, F, H, I) 0.75–1.125 M sucrose gradients that partially resolve Golgi and IC membranes at 15°C. On these shallow gradients, ER membranes pellet and are not included above. Gradient fractions were immunoblotted to detect the indicated proteins, and the amounts of each protein were quantitated by densitometry and plotted on the ordinates above. The 41-kD and 34-kD isoforms of syntaxin 5 are presented separately in D, E, and F. Fractions are displayed from least dense to most dense.