Anti cxcl8

Kidney tissues in each group were fixed in 4% paraformaldehyde and blocked in paraffin. Thereafter, the sections were stained in hematoxylin for 5 min and eosin for 3 min. Images of HE sections were observed under an inverted microscope (Leica, Germany). 5 fields were randomly determined from each section. The kidney damage was evaluated according to the loss of brush borders (0–3), tubular vascularization (0–3) and inflammatory cell infiltration (0–3) [Zhou, 2014 #48]. All sections were independently assessed by an investigator in a blinded manner. IHC was performed as previously described [33 (link)]. To be more specific, the sections were washed by PBS and blocked in 3% H₂O₂ for 10 min. and were then incubated with goat serum for 15 min and washed. Subsequently, the sections were incubated with anti-CXCL8 (1:500, Abcam, Cambridge, UK) overnight at 4 °C followed by being incubated with streptavidin peroxidase-conjugated second antibody (Zhongshan Golden Bridge, Beijing, China) for 1 h at 37 °C. Next, the sections were stained with 3,3′-diaminobenzidine (DAB, Zhongshan Golden Bridge) for 6 min and then rinsed and stained in hematoxylin for 30 s. The sections were dehydrated in gradient alcohol and sealed in neutral resins. The integral optical density (IOD) was calculated with Image Pro plus 6.0 software (Microsoft Media Cybernetics, Bethesda, MD, USA).