Skip to main content

Advertisement

Fig. 5 | Cell Communication and Signaling

Fig. 5

From: FAF1 mediates necrosis through JNK1-mediated mitochondrial dysfunction leading to retinal degeneration in the ganglion cell layer upon ischemic insult

Fig. 5

FAF1-mediated JNK1 activation triggers mitochondrial dysfunction upon ischemic stress. a Faf1+/+MEFs were treated with oxygen glucose deprivation for the indicated times and then fractionated into cytoplasmic and mitochondrial fractions. The fractions were analyzed by immunoblotting with anti-P-JNK, JNK1, anti-COX IV (mitochondrial marker) and anti-β-actin (cytoplasmic marker) antibodies. b Faf1+/+ and Faf1gt/gt MEFs were treated with oxygen glucose deprivation for 2 h and then fractionated into cytoplasmic and mitochondrial fractions. The fractions were analyzed by immunoblotting with anti-P-JNK, JNK1, anti-COX IV and anti-β-actin antibodies. c Upper panel: Faf1+/+ and Faf1gt/gt MEFs were transfected with DsRed-Mito plasmid for 48 h. Subsequently, the cells were treated with oxygen glucose deprivation in the presence or absence of 20 μM SP600125 for 2 h. Fluorescence images show the alterations in mitochondrial morphology. Scale bar = 20 μm. Bottom panel: the graph shows the quantification of mitochondrial fragmentation (n = 3). d Faf1+/+ and Faf1gt/gt MEFs were untreated or treated with oxygen glucose deprivation in the presence or absence of 20 μM SP600125 for 5 h. Mitochondrial superoxide was stained using 10 μM MitoSOX reagent. The population of MitoSOX-positive cells was detected by flow cytometry. The graph shows the results of the quantitative analysis of MitoSOX-positive cells (n = 3). e Faf1+/+ and Faf1gt/gt MEFs were untreated or treated with oxygen glucose deprivation in the presence or absence of 20 μM SP600125 for 5 h, and the mitochondrial potential was measured using a Muse analyzer. The graph was obtained from a quantitative analysis of depolarized cells (n = 3). The data (c - e) are expressed as the mean ± S.E.M. of three independent experiments. Statistical comparisons were evaluated using ANOVA followed by Tukey’s HSD (c - e) post hoc analysis. ***P < 0.001, **P < 0.01, and *P < 0.05

Back to article page