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. 2014 Mar 25;15(5):540–547. doi: 10.1002/embr.201337948

Figure 3. Somatic or axonal LIGHT signaling elicits opposite responses in motoneurons.

Figure 3

A Schematic representation of the microfluidic device.

B Motoneurons are plated into the somatic compartment and they extend their axons into the axonal compartment through 750-μm-long microgrooves.

C, D Hb9::GFP motoneurons were cultured for 120 h and immunostained using SMI312 (C) and MAP2(D). Scale bar, 50 μm.

E, F After 120 h of culture, sLIGHT (100 ng/ml) was added either in the somatic or in the axonal compartment, or both. Axonal length (E) and branching (F) were determined 48 h later.

G The retrograde transport of Alexa555-conjugated WGA allows tracing of motoneurons whose axons have crossed microchannels (white arrows).

H The survival of motoneurons whose axons have reached the axonal compartment is determined 48 h later by counting the number of GFP- and WGA-positive cells. Scale bar, 100 μm.

Data information: Histograms show mean values ± SEM in three independent experiments, ANOVA with Tukey–Kramer’s post hoc test; *P < 0.05, ***P < 0.001