Fig. 4
Reactive astrocytes expressed fibronectin both in vitro and in vivo. a Sagittal sections of chronically injured spinal cord. Asterisk indicates the lesion epicenter. Fibronectin, Laminin, white. Scale bar, 500 μm. b Sagittal section of naïve spinal cord. Magnification of the inset is shown in d. GFAP, red. c Sagittal section of injured spinal cord at 7 days post-injury. Magnification of the inset is shown in e. GFAP, red. Asterisk indicates the lesion epicenter. Scale bar, 500 μm. d, e GFAP-positive astrocytes (marked by white arrow-heads) were isolated marginally (surrounded area by white dots) by laser-captured microdissection (LMD). GFAP, red; Hoechst, blue. Scale bar, 20 μm. f Both NAs and RAs were isolated from spinal cord by LMD in vivo. Purple dots indicate the cutting line of the peri-lesional area. The mRNA expression of fibronectin and laminin, ligands of β1 integrin receptor, was analyzed by qPCR. Error bar indicates mean ± SEM. Star indicates statistical significance (p < 0.05). Wilcoxon’s rank-sum test. n = 3 per each group, triplicate. Fibronectin: F = 1.3 × 10−6. Laminin: F = 0.0385. g Both NAs and RAs were collected from primary cultures in vitro. The mRNA expression of fibronectin and laminin was analyzed by qPCR. Error bar indicates mean ± SEM. Star indicates statistical significance (p < 0.05). Wilcoxon’s rank-sum test. n = 3 per each group, triplicate. Fibronectin: F = 0.0432. Laminin: F = 0.0002