The Journal of Neuroscience, September 13, 2006, ():
Activated c-Jun N-Terminal Kinase Is Required for Axon Formation
J. Neurosci. Oliva et al.
26: 9462
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1. SP6000125-treatment abolished phospho-JNK immunoreactivity, whereas cellular stress led to strong phospho-JNK immunostaining throughout the entire neuron. Left panels, Control condition. Center panels, Cultures treated for 90 min with SP600125 showed an almost complete absence of phospho-JNK immunostaining. Right panels, Cultures treated for 5 min with 400 mM sorbitol. This standard paradigm for hypo-osmotic stress-activation of JNK resulted in robust phospho-JNK immunostaining throughout the neurons. These results demonstrate that the pool of available JNK expressed throughout the all the neurites and soma of stage 3 neurons can be activated, even though JNK is not activated in the minor processes under normal conditions. Arrows indicate axons. Scale bars: 20 μm.
- supplemental material
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Supplemental Figure 2. (a) Representative neuron at 4 weeks in culture expressing YFP-JBD (green, left panel). This neuron had no axon, but axonal processes from neurons separately transfected with CFP (red) and co-plated can be seen (center panel). Right panel is a phase image of the culture, demonstrating that the culture itself was healthy. (b) Quantitative analysis of axon formation in neurons transfected to express YFP-JBD (white bars) or soluble cyan fluorescent protein (CFP; black bars).
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Supplemental Figure 3. Phosphorylated ATF-2 is enriched in axons. Representative stage 3 neuron transfected with YFP (a) and immunohistochemically processed for dually phosphorylated ATF-2 (b; phospho-ATF-2). Phospho-ATF-2 was enriched in the axon in a distally-directed gradient that tapered off near the growth cone (c), similar to phospho-JNK. Arrow indicates the axon. Scale bar: 25 μm.
