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. 2018 Jun 6;38(23):5399–5414. doi: 10.1523/JNEUROSCI.3214-17.2018

Figure 5.

Figure 5.

Pretreatment of aggrecan spots with ISP-treated CM or rCatB enhances axon crossings through the aggrecan gradient. A, EnzChek protease assay of recombinant CatB ± recombinant CSTB (rCSTB) (n = 6; F(4,5) = 11.32, p = 0.0101, ANOVA). B, Aggrecan spot degradation with CM from vehicle, 2.5 μm ISP, or S-ISP-treated DRGs, or media-only, 0.1 U/ml ChABC, 50 ng/ml recombinant CatB, 10 ng/ml recombinant CSTB alone or in combination with vehicle, or 2.5 μm ISP-treated DRG CM (n = 106, 157, 141, 136, 21, 45, 11, 63, 47; F(12,155) = 30.35, media-only vs ChABC, p = 0.0019; media-only vs ISP, p = 0.0001; media-only vs rCatB, p = 0.0001; ISP vs ISP+rCSTB, p = 0.0001; ANOVA). C, D, Axon crossings normalized by total neurons present on aggrecan spots pretreated with CM collected from vehicle, 2.5 μm ISP or SISP, or 0.1 U/ml ChABC, recombinant CatB (0.5 μg/ml) with or without recombinant CSTB (50 μg/ml) with representative images. Scale bar, 50 μm. Dotted lines indicate CSPG border. Inset, Axon crossing CSPG gradient (n = 11, 20, 12, 12, 12, 17, 30, 29; F(7,32) = 9.719, media vs ChABC, p = 0.0018; S-ISP vs rCatB, p = 0.0207; ISP CM vs ChABC, not significant, p = 0.5606; ISP vs ISP+rCSTB, p = 0.0402; ANOVA). Lines indicate median. Boxes represent quartiles. Whiskers indicate range. *p < 0.05, **p < 0.01, ***p < 0.001.