Fig. 1

JNK1 contributes to necrosis upon ischemic stress induced by oxygen-glucose deprivation in MEFs. a MEFs were treated with oxygen glucose deprivation for the indicated times and/or 1 μM staurosporine (STS) for 12 h. The type of cell death was determined by flow cytometry using double staining with annexin V and PI. b The graph shows the quantitative results of the flow cytometry analysis. c and d MEFs were treated with oxygen glucose deprivation or 1 μM STS for the indicated times (n = 3). Caspase-3 activity was measured using fluorometric assay (n = 3) (c) and western blot analysis (d). e MEFs were untreated or treated with oxygen glucose deprivation for 8 h in the presence or absence of zVAD-fmk (50 μM), and cell death was determined by measuring PI uptake using a flow cytometer (n = 3). f MEFs were pretreated with Nec-1 (50 μM), DPQ (30 μM), or SP600125 (20 μM) for 30 min and then with oxygen glucose deprivation for 8 h in the presence of the individual compounds. Cell death was detected by flow cytometry (n = 3). g Left panel: Jnk1+/+ and Jnk1−/− MEFs were untreated or treated with oxygen glucose deprivation for 8 h. Cell death was detected using flow cytometry (n = 3). Right panel: representative immunoblots show the JNK1, FAF1 and β-actin expression levels. The data (b, c, e-g) are expressed as the mean ± S.E.M. of three independent experiments. Statistical comparisons were evaluated using ANOVA followed by Dunnett’s T3 (b, c, and e) and Tukey’s HSD (f and g) post hoc analysis. ***p < 0.001, **P < 0.01, and *P < 0.05