Figure 2.

Representative western blots of proteins in exosomes derived from Wnt3a treated microglia. Primary microglia (2500000 cells per treatment) were treated with or without Wnt3a (10 nM) for 8 hours in SFM (Control: ‘Con’ = without Wnt3a) (A). Cell culture medium was collected, centrifuged at 10000xg for 10 minutes, concentrated using 3kDa centrifugal devices and exosomes were isolated at 100000xg. Smaller membrane vesicles were extracted at 200000xg for 16 hours. Proteins were separated by SDS-PAGE and subjected to western blotting for Wnt3a, β-actin and Alix. Primary microglia (12500000 cells per treatment) were treated with or without Wnt3a (10 nM) for 36 hours in SFM (Control: ‘Con’ = without Wnt3a) (B). Cell culture medium was collected, centrifuged at 10000xg for 10 minutes, concentrated using 3kDa centrifugal devices and exosomes were isolated at 100000xg. Proteins were separated by SDS-PAGE and subjected to western blotting for Wnt3a and Alix.

Hooper et al. BMC Neuroscience 2012 13:144   doi:10.1186/1471-2202-13-144
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