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. Author manuscript; available in PMC: 2011 Jun 22.
Published in final edited form as: Neurobiol Dis. 2008 Jul 8;32(1):105–115. doi: 10.1016/j.nbd.2008.06.015

Figure 5. Myelin production by neuropathic explant cultures is enhanced by EC137.

Figure 5

(A) Schematic of the treatment paradigm for DRGs from Wt and C22 mice with EC137. The black line indicates the time-scale for days in vitro (DIV). The lines in green, bold and dashed, represent the expression profiles of myelin proteins in DRGs from Wt and C22 mice respectively. The blue line indicates the time-scale (h) for EC137 treatment. Starting on DIV21, a pulse treatment of EC137 (50 nM) (green arrows) was added for 48 h, followed by 48 h washout (wo, black arrows). This sequence was repeated and a second wash out (16 h) was followed by analyses of the samples (arrow head). (B) DRG explant cultures from Wt (top panel) and C22 neuropathic (bottom panel) mice, under myelinating conditions, were treated with DMSO (control) or EC137 (50 nM) for a total of 96 h as described (A), and stained with an anti-MBP antibody. Insets show the outlined regions at 3X magnification. Hoechst dye was used to stain the nuclei. Scale bar, 40 μm. (C) The lengths of the myelin internodes (n is at least 100 for each condition) were measured in explant cultures from Wt and C22 mice treated with DMSO or EC137 (50nM), using Spot Advanced software. *p<0.05, ***p< 0.001. Error bars show SEM. (D) DRG explants from Wt and C22 mice were treated as described (A) and whole protein lysates (40 μg/lane) were analyzed for the levels of myelin proteins MAG, P0 and MBP, and of HSP70 from at least three independent experiments. Arrows on the MBP blots indicate the 21.5, 18.5, 17 and 14 kDa isoforms. GAPDH serves as a loading control. Molecular mass in kDa.