Fig. 4

Silencing FANCC exacerbates SCI-induced pyroptosis and NLRP3-depended neuroinflammation. A Western blotting of FANCC, p-p38, p38, ASC, GSDMD and GSDMD-N protein levels at 3 dpi in NC-sham, NC-SCI and KD-SCI mice. B Bar graph showing a quantitative analysis of FANCC expression; n = 3. C Bar graph showing the ratio of p-p38/p38; n = 3. D Densitometric analysis of ASC expression. E Densitometric analysis of GSDMD expression. F Densitometric analysis of GSDMD-N expression. G Double immunofluorescence of Caspase-1 (green) and NLRP3 (red), obtained from longitudinal sections centered around central canal at 3 dpi in Sham, SCI and KD-SCI mice. Scale bar = 100 μm. H Double immunofluorescence labeling of microglia for IBA-1(green) and iNOS (red), obtained from longitudinal Sects. 1 mm caudal to the lesion site at 3 dpi in Sham, SCI and KD-SCI mice. As shown in the KD-SCI mice, iNOS was increased in the microglia. Scale bar = 100 μm. I, J ELISAs performed for the IL-1β and IL-18 expressions in injured tissue obtained at 3 dpi, showing significantly increased levels of IL-1β and IL-18 in KD-SCI group; n = 5. The error bars represent the SD. *p < 0.05 vs. NC-sham group, #p < 0.05 vs. NC-SCI group by one-way ANOVA followed by Tukey’s post hoc analysis (*p < 0.05, **p < 0.01, and ***p < 0.001). NC-sham mice were performed laminectomy only, NC-SCI mice were performed spinal cord contusion, KD-SCI mice were transfected with KD-FANCC before SCI