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Figure 1


Fig. 1. The exocyst components Sec8 and Sec6 are present in OLs and myelin. (A) OL lineage cells, myelin, brain homogenate and astrocytes were analyzed by immunoblot (10 µg protein/lane). The levels of Sec8 and Sec6 were normalized to actin and quantified for n=3 independent OL cultures. (B-E) Sec8 (green) colocalization with (B) TGN38 (red) and (C) LAMP2 (red) in OL processes was assessed by confocal microscopy and ratiometric particle analysis for (D) 434 and (E) 560 randomly chosen puncta in eight representative OLs, two independent cultures per condition. (B) The arrows indicate colocalization between TGN38 and Sec8, shown at a higher magnification in the inset. (F) Sec8 (green) is detectable along MBP-positive (red) myelinated axons in mouse spinal cords sectioned longitudinally (left panel) or transversally (right panel). No immunofluorescence was detected when cells and/or sections were incubated with pre-immune IgGs and secondary Abs (data not shown). EP, early OL progenitors; LP, late OL progenitors; OL, mature OLs; BH, whole-brain homogenate; My, myelin; As, astrocytes. Bars, 10 µm (B,C), 2 µm (F).





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