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. Author manuscript; available in PMC: 2010 Jun 1.
Published in final edited form as: Nat Neurosci. 2009 Nov 15;12(12):1534–1541. doi: 10.1038/nn.2446

Figure 6. Jedi-1 and MEGF10 expressed in HEK 293 cells enable binding to dead neurons.

Figure 6

HEK 293 cells were transfected with GFP-tagged Jedi-1 or MEGF10 and their expression on the cell surface analyzed by biotinylation of the surface proteins followed by precipitation with avidin beads and Western blotting with anti-GFP (a). The biotin reagent was not added to some cells (no biotin) to confirm the specificity of the pull down. The expression of Jedi::GFP and MEGF10::GFP in total cell lysates is shown in the lower panel. (b) Jedi-1 and/or MEGF10 transfected cells were incubated at 4°C with the indicated number of neuronal corpses, induced to undergo apoptosis by NGF withdrawal for 24 hrs and labeled with propidium iodide. After washing off the unbound dead cells, the cultures were fixed with 10% formalin. The percentage of GFP+ cells with at least one PI+ corpse bound is shown (1 representative experiment of 3 is depicted). The lower panel shows the mean ± S.E.M. percent binding at the highest concentration of neurons (*, p < 0.01 relative to GFP transfected cells; n=4).