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. 2016 Nov 15;6:37050. doi: 10.1038/srep37050

Figure 3. Calpain-mediated cleavage of CRMP2 during acute axonal degeneration in vivo and after scratch in vitro.

Figure 3

(a,b) Representative Western blots of CRMP2 proximal (a) and distal (b) to the crush site in uncrushed native optic nerves (0 h) and in optic nerves at 6 h after crush. Below, quantification of cleaved CRMP2 band intensity relative to GAPDH. 4 optic nerves per group. Error bars represent standard error of the mean (SEM). **P < 0.01, *** P < 0.001 by independent samples t-test. (c,d) Representative Western blots of CRMP2 proximal (c) and distal (d) to the crush site in uncrushed native optic nerves (control), in optic nerves at 6 h after crush pretreated with 7% DMSO (crush + DMSO), and in optic nerves at 6 h after crush pretreated with 10 mM calpeptin in 7% DMSO (crush + cal). Cal = Calpeptin. Quantifications of cleaved CRMP2 band intensity relative to GAPDH are shown below. Four optic nerves are included in each group. Error bars represent the standard error of the mean (SEM). **P < 0.01 by one-way ANOVA and Dunnett’s test. (e,f) Representative Western blots of spectrin (e) and CRMP2 (f) in unscratched primary cortical neuron cultures pretreated with 0.1% DMSO (control), scratched cultures pretreated with 0.1% DMSO (scra + DMSO), and scratched cultures pretreated with calpeptin in 0.1% DMSO (scra + cal). Cal = calpeptin. Sca = scratch. Below, band intensities of 145 kDa cleaved spectrin (e) and cleaved CRMP2 (f) were quantified and normalized to GAPDH. 3 independent cultures per group. Error bars represent the standard error of the mean (SEM). *P <  < 0.05, **P < 0.01 by one-way ANOVA and Dunnett’s test.