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

Figure 4.

Figure 4.

ISP promotes secretion of CatB. A, ISP-dependent aggrecan spot degradation is not dependent on transcription or translation, as shown by anisomycin (20 μg/ml), cycloheximide (20 μg/ml), or α-amanitin (20 μg/ml) treatment of DRGs (n = 103, 91, 54, 50, 73, 57, 72, 60; F(7,113) = 2.6, not significant; ISP vs anisomycin+ISP, p = 0.3855; ISP vs cycloheximide+ISP, p = 0.490; ISP vs α-amanatin, p = 0.6309; ANOVA). B, Inhibiting exocytosis with a high concentration of Exo1 (10 μg/ml) rescues GAG chain degradation by ISP (n = 115, 187, 160, 76; F(3109) = 35.18, DMSO vs ISP, p = 0.0003; ISP vs Exo1+ISP, p = 0.0108; ANOVA). C, Western blot of CatB and CSTB from DRG CM collected after 2 or 4 DIV treated with vehicle control, 2.5 μm ISP, or S-ISP; n = 5. The same blot was stained with total protein dye, Coomassie Blue. D, Western blot of CatB, CSTB, or GAPDH from DRG lysate treated with vehicle control, 2.5 μm ISP, or S-ISP for 4 DIV; n = 3. DRGs stained with Tuj1 and CSTB (E) or CatB (F). Scale bar, 50 μm. Lines indicate median. Boxes represent quartiles. Whiskers indicate range. **p < 0.01, ***p < 0.001.