Fig. 3
Knockdown of HSPA8 inhibits NLRP3 inflammasome activation and phosphorylation of NF-κB in SCII rats. A Immunofluorescence of GFAP in spinal cord tissues after SCII was performed. Scale bar = 50 μm. B Quantification of GFAP-positive cells after SCII was analyzed. C Relative protein levels of NLRP3, ASC, and p20/pro-caspase-1 in spinal cord tissues of SCII rats were detected after injection of LV-sh-HSPA8 via western blot assay. D The contents of IL-1β and IL-18 in the spinal cord tissue homogenate were detected by ELISA assay. E Relative protein levels of total NF-κB P65 and p-NF-κB P65 in spinal cord tissues of SCII rats were detected after injection LV-sh-HSPA8 via western blot assay. F The intracellular location of the NF-κB P65 in spinal cord tissues of SCII rats was observed by immunofluorescence staining. Scale bar = 50 μm. G The DNA-binding activity of NF-κB in spinal cord tissues of SCII rats was revealed by EMSA. Data were shown as means ± SD of three independent experiments. N = 6 rats per group. Statistical analysis was performed by one-way ANOVA followed by Turkey’s for multiple comparisons. *p < 0.05 vs. sham. # p < 0.05 vs. SCII + LV-sh-NC. NLRP3, nod-like receptor pyrin domain-containing 3; ASC, the apoptosis-associated speck-like protein containing CARD; SCII, spinal cord ischemia–reperfusion injury; IL-1β/18, interleukin-1β/18; NF-κB, nuclear factor-kappa B