A00533

After rats were anesthetized and rapidly perfused with 4% paraformaldehyde, brain tissues were removed and placed in 4% paraformaldehyde at 4°C overnight for post-xation. The brain tissues were placed in 20% sucrose for dehydration for 2 days and then replaced with 30% sucrose for dehydration for another 2 days. The brain tissues were cryosectioned to a thickness of 20 μm and further immune-stained with uorescence. Brie y, sections were incubated with 1% BSA at room temperature, and next incubated overnight at 4°C with the following antibodies: CXCL1 antibody (A00533, rabbit, 1:50, Boster, Wuhan, Hubei, China), CXCR2 antibody (BA0732-2, rabbit, 1:50, Boster, Wuhan, Hubei, China), GFAP antibody (astrocyte marker, MAB360, mouse, 1:1000, Millipore, Billerica, MA, USA), IBA-1 antibody (microglial cell marker, ab5076, goat, 1:500, abcam, Boston, USA) and NeuN antibody (neuron marker, MAB377, mouse , 1:1,000, Millipore, Billerica, MA, USA). The next day, the sections were incubated with Cy3-linked secondary antibody or Alexa 488-linked secondary antibody (1:1000, Jackson ImmunoResearch, West Grove, PA) for 2 h at room temperature. The stained sections were rinsed three times on a shaker for 15 min each, then air dried and sealed on slides. The stained slides were observed by a Nikon uorescence microscope and images were captured by CCD Spot.