Figure 5

Peli1 deficiency inhibited microglia-mediated neuron death. (a) Scheme of conditioned media (CM) and cell death assay by using WT and/or Peli1-KO microglia or astrocytes. (b,c) CMs were harvested from WT and Peli1-KO microglia that were stimulated with LPS (100 ng/ml) for 24 h, and then used for the test of viability of Neuro2A cells (b) or primary neurons (c). For sequential CM assay, CMs harvested from microglia were cultured with WT and Peli1-KO astrocytes for 24 h. Then, CMs were harvested and tested for effect on viability of Neuro2A cells. Neuro2A cells that treated with TNF (20 ng/ml) plus cycloheximide (CHX) were used as a positive control. (d) Effect of Peli1 knockdown in Neuro2A cells on sensitivity to TNF (20 ng/ml) plus CHX induced apoptosis assessed by flow cytometry assay, showing the frequencies of Annexin V⁺ cells in bar graph. Data with error bars represent mean ± SD. Each panel is representative of at least three independent experiment. *P < 0.05, **P < 0.01 as determined by one-way ANOVA with a Tukey’s post test (b–d).