Blagoveschenskaya et al. Vol. 145, No. 7, June 28, 1999. Pages 1419–1433.
An incorrect graphic appears in the legend for Fig. 1. The corrected sentence appears below in bold.
Figure 1. A two-step subcellular fractionation procedure for separation of late endosomes and DCG. (A) Distribution of intracellular compartments from PC12 cells on 1–16% Ficoll gradients. PC12 cells expressing ssHRPP-selectin were loaded with 3H-Dopamine, or labeled with 125I-Trn or 125I-EGF as described in Materials and Methods. Cells were homogenized in HB and the PNS was centrifuged on 1–16% Ficoll gradients and fractionated. The early/recycling endosomes are shown by the distribution of 125I-Trn endocytosed for 60 min at 37°C (▵). Late endosomes are shown by the distribution of 125I-EGF internalized for 20 min at 37°C (▪). The distribution of NAGA activity (OD420 nm; ○), HRP activity (OD450 nm; •), and 3H-Dopamine radioactivity (filled plus signs) along 1–16% Ficoll gradients are shown. (B) Separation of DCG and late endosomes on a secondary 0.9–1.85 M sucrose gradient. Fractions 14–20 from the 1–16% Ficoll gradients were pooled, diluted with HB, and recentrifuged on 0.9–1.85 M sucrose gradients to equilibrium. The distributions of NAGA activity (○), HRP activity (•), 3H-Dopamine radioactivity (filled plus signs), 125I-EGF internalized for 20 min at 37°C (▪) and 125I-Trn (▵) after centrifugation on this gradient are shown.
