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. Author manuscript; available in PMC: 2013 Apr 1.
Published in final edited form as: Nat Protoc. 2012 Sep 6;7(10):1774–1782. doi: 10.1038/nprot.2012.100

Figure 4.

Figure 4

Myelination of hippocampal neurons expressing YFP. (a) A YFP-expressing hippocampal neuron at 21 DIV in regular culture. The neuron was transfected with YFP (green; left) at 18 DIV, using the Ca2+-phosphate method32. The transmitted light image is on the right. (b–d) The soma (white asterisk) and proximal axon (white arrowheads) (b) of a YFP-expressing hippocampal neuron in myelin coculture at 30 DIV; the hippocampal neurons were transfected at 7 DIV using Lipofectamine 2000, before oligodendrocytes were seeded (b–d). A myelin segment, revealed by the rat monoclonal anti-myelin basic protein (MBP) antibody (1:200 dilution; Chemicon) (red), formed along the proximal axon. YFP is in green. (c,d) YFP-positive axons of transfected hippocampal neurons were myelinated revealed by MBP staining (white arrows)30. Scale bars: a, 100 μm; b, 50 μm; c,d, 25 μm.