Bs 6655r

On postoperative day 7, the L4–L6 spinal cord was harvested from rats (n = 3 for SCI group), embedded in paraffin, and cut into 20-μm-thick sections. These sections (n = 3 for each sample) were dewaxed, dehydrated by gradient alcohol, and repaired by antigen. Next, the sections were blocked with endogenous peroxidase and 10% donkey serum and then incubated with the following primary antibodies: NLRP3 (1:200; bs-6655R, Bioss), GFAP (1:500; GB12096, Servicebio), IBA-1(1:500; GB12105, Servicebio), and NeuN (1:100; GB13138-1, Servicebio) overnight at 4°C. After the primary antibody incubation, the sections were incubated with the corresponding secondary antibodies conjugated with CY3 and FITC for 1 h in dark conditions at 37°C. Finally, the dorsal horns were observed and photographed under a fluorescence microscope (Olympus, Japan).

The paraffin-embedded renal cortices Sec. (3 μm) were incubated with primary antibodies against podocin (NPHS2, Abcam, # ab50339, 1:200) and NLRP3 (NALP3, BIOSS, #bs-6655R, 1:200) overnight at 4 °C. Ten fields of view (400 × magnification) per slide were randomly selected for analysis and the percentage of positively-stained area over the whole glomerular area was calculated using Image J Software (National Institutes of Health).