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Fig. 2. Isolation of large and small prominin-1-containing membrane particles from the embryonic ventricular fluid. (a) Ventricular fluid of E10.5-E11 mouse embryos was subjected to differential centrifugation for 5 minutes at 300 g (P1), 20 minutes at 1,200 g (P2), 30 minutes at 10,000 g (P3) and 1 hour at 110,000 g (P4). The entire P1-P4 pellets were analysed by immunoblotting for prominin-1. (b,c) Immunofluorescence of prominin-1 of P2 and P4 particles obtained by differential centrifugation of E13 ventricular fluid for 15 minutes at 10,000 g (b) and 1 hour at 200,000 g (c), respectively. The inset in b shows a P2 particle at higher magnification, which is very similar to the one shown in the inset of Fig. 1f. (d-f) Negative staining EM of prominin-1 immunogold labelled P4 (d) and P2 (e,f) particles obtained by differential centrifugation of E11.5 ventricular fluid as in a.





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