Fig. 6

Assessment of iNOS expression in cultured BMDMs following CSPG stimulation; MMP 5-plex assay and ELISA analyses of conditioned media from cultured BMDMs. Cultured BMDMs from wild-type and EXTL2^−/− mice were stained for iNOS and nuclear yellow (NY). Representative images for unstimulated and CSPG-stimulated cells are shown in a. Scale bar = 100 μm. b Comparison of number of iNOS⁺ cells between wild-type and EXTL2^−/− cells, unstimulated or with CSPG stimulation. n = 4 wells per condition. Two-way ANOVA with Sidak’s multiple comparisons test. c-g Fold change of MMP2, MMP8, MMP3, pro-MMP9, and MMP12 detected in conditioned media from both wild-type and EXTL2^−/− BMDMs, unstimulated and CSPG stimulated, respectively. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Raw quantities of MMPs in pg/mL ranged from 37.1 to 58.1 for MMP2, 0.1 to 10.3 for MMP3, 305.7 to 490.0 for MMP8, 157.6 to 6265.0 for pro-MMP9, and 3936.5 to 11,558.3 for MMP12. Fold change is calculated over unstimulated cells. h TNFα levels in BMDM conditioned media as determined by ELISA. Two-way ANOVA with Sidak’s multiple comparisons test. i Representative images of TNFα and CD68 staining in day 14 lysolecithin-injured spinal cord tissue. Scale bar = 200 μm. j Analysis of absolute TNFα immunoreactive area within lesions at day 14 in wild-type and EXTL2^−/− spinal cords. All graphs are mean with SEM, except for panel j (mean with standard deviation). Two-tailed, unpaired Student’s t test. *p < 0.05, **p < 0.01