e13 5 cd1 mouse embryos  (Charles River Laboratories)

Journal: Nature neuroscience

Article Title: Glial Precursors Clear Sensory Neuron Corpses during Development via Jedi-1, an Engulfment Receptor

doi: 10.1038/nn.2446

Figure Lengend Snippet: DRG from E13.5 mice were dissociated and grown in the presence of NGF for 2 days. The glial cells were then transfected with plasmids expressing MEGF10::GFP; Jedi-1::Flag, or meGFP and NGF removed from the cultures. The cells were fixed after 2 days without NGF and immunolabeled with antibodies to GFP or Flag. (a and b) Z-axis optical stacks of confocal images of glial cells expressing MEGF10::GFP (M10::GFP; a, c and d), Jedi-1::Flag (Jd::Flag; b, c and d), or meGFP (c and d) were acquired and the numbers of transfected cells containing at least one ingested nuclear remnant [condensed TO-PRO3 staining] was quantified. Arrows indicate the location of engulfed apoptotic nuclei in these cells. Arrowheads indicate the long processes in Jedi-1::Flag expressing cells. In (c) the error bars = Mean ± s.d. P=0.0002, one-way ANOVA. (d) The number of engulfed nuclei per engulfing cell was also determined and expressed as the percent of transfected cells containing the indicated number of apoptotic nuclei (open bar: meGFP; black bar: MEGF10-GFP; gray bar: Jedi-Flag). Based on a chi-squared analysis, there was a significant difference between all 3 groups of cells (expressing Jedi-1, MEGF10 and meGFP. p<0.0001). Scale bars, 20 µm.

Article Snippet: DRG neurons from E13.5 CD1 mouse embryos were isolated as described above, plated on collagen coated coverslips and grown in UltraCULTURE media with 50 ng/ml NGF (Harlan).

Techniques: Transfection, Expressing, Immunolabeling, Staining