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Fig. 3. Netrin-1-mediated DCC-dependent commissural axon outgrowth requires lipid raft integrity. E13 dorsal spinal cord explants were cultured for 18 hours (A,B) or 40 hours (C) in collagen gel either without a netrin-1 source (–), or with purified netrin-1 (Net-p) or next to ventral spinal cord explants (FP). As described in the Materials and Methods, explants were either left untreated (–), treated for 1 hour with 2 U/ml of CO (CO1), 1.5 hours with 1 U/ml of CO (CO2) or 12 minutes with 10 mM MßCD. (B) Quantification of A. The total number of explants that were quantified from four distinct experiments varies from 10 to 15 per tested condition. Values shown are means ± s.e.m. Bars, 170 µm. (D) Quantification (as in B) of the effect of depletion-repletion in cholesterol on E13 dorsal spinal cord explants. Explants were incubated with MßCD as in A and further treated with 1 mM cholesterol and then cultured for 18 hours in collagen gel. Values shown are percentages of netrin-1 treated explants (100%) ± s.e.m.
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