Fig. 4

Conditioned knockout of microglial Fascin-1 restricts tissue softening after SCI. (A) Construction diagram of Fascin-1 CKO mice. (B) Sagittal immunofluorescence images of CX3CR1 (red) and Fascin-1 (green) from Fascin-1^fl/fl and Fascin-1 CKO mice at 14 dpi. Scale bar: 200 μm. (C) Quantitative analysis of the density of Fascin-1⁺CX3CR1⁺ microglia. (D, E) Sagittal immunofluorescence images for microglia (CX3CR1, red) and astrocyte (GFAP, green) from Fascin-1^fl/fl and Fascin-1 CKO mice at 7 (D) and 14 dpi (E). Region a represents the GFAP⁻ lesion core, and region b represents the GFAP⁺ penumbra region. Scale bar: 100 μm. (F, G) Comparison of the elastic properties of lesion core (a, F) and penumbra region (b, G) in two groups of mice as described above in (D) and (E). (H, J) Sagittal immunofluorescence images for the astrocyte marker GFAP (green, H) and vimentin (green, J) in Fascin-1^fl/fl and Fascin-1 CKO groups before SCI and at 7 and14 dpi. The nuclei were marked with DAPI (blue). Scale bar: 100 μm. (I, K) Percentage quantification of GFAP⁺ area (I) and vimentin⁺ area (K). The lesion core was marked with the asterisks. n = 3 animals in (C), (F), (G), (I), and (K). ns, no signifcance; ****P < 0.0001 by Student’s t test in (C). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by two-way ANOVA in (F), (G), (I), and (K)