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Total RNA was isolated as described [71], fractionated on an agarose/formaldehyde denaturing gel, and immobilized onto a Hybond-N+ membrane (Amersham).

Sucrose gradients (5 20%) were poured as previously described [37] and fractionated from the top by hand.

Glycolipids were extracted from cells as described elsewhere, 9 fractionated by thin‐layer chromatography on high‐performance thin‐layer chromatography plates (Merck, Darmstadt, Germany) and visualized with orcinol H2SO4.

For co-IP of NPRA with plasma membranes, early endosomes, lysosomes and Res, cells were fractionated as described, by subcellular fractionation.

Cultures of BL21 DE3)/pX12345 at different induction time points were harvested and fractionated as described in "Materials and methods" section.

Brain extracts were fractionated as described [53].

Cells were fractionated as described previously [12] with modifications.

Cells were fractionated as described [47] with minor modifications.

Mouse brain homogenates were fractionated as described [19].

Cells were grown in 100 mm tissue culture dishes as described above and cell fractionated essentially as described by Blobe et al. [58].

Lysates of HeLa Tet-on cells (2×10-cm plates) transfected with siRNAs for 48 hrs were fractionated as described [44] with one significant modification.