Df6421

Renal tissue sections including six patients of FSGS and three kidney para‐cancerous samples were obtained from the Second Affiliated Hospital of Guangxi Medical University. Ethical approval for this study was obtained from The Human Research Ethics Committee of the Second Affiliated Hospital of Guangxi Medical University. All patients provided written and verbal informed consent. Then tissue sections were incubated overnight at 4 °C with rabbit monoclonal antibodies against ALB (16475‐1‐AP; dilution 1 : 100; ProteinTech, Rosemont, IL, USA), APOE (AF5178; dilution 1 : 150; Affinity, Cincinnati, OH, USA), CLU (DF6421; dilution 1 : 100; Affinity), CST3 (DF6457; dilution 1 : 120; Affinity), SERPINA1 (DF6182; dilution 1 : 100; Affinity) and SPP1 (BS‐0026R; dilution 1 : 150; BIOSS, Woburn, MA, USA). This was followed by incubation with secondary antibodies at room temperature for 20 min. Tissue sections were dyed with hematoxylin and then counterstained, dehydrated and mounted. IHC results were analyzed using imagej (NIH, Bethesda, MD, USA).

EVT-1 cells were treated with LPS for 24 h, whole lysates were prepared by direct lysis in RIPA buffer with PMSF (Beyotime, Beijing, China). Electrophoresis processes were performed as previously described [18 ]. Primary antibodies of CLU (DF6421, Affinity, Cincinnati, OH, USA) and GAPDH (AF7021, Affinity, Cincinnati, OH, USA) were used in this analysis. Antigens were revealed using a chemiluminescence assay (Affinity, Cincinnati, OH, USA). Quantification of bands was achieved by densitometry using the Chemiluminescence Image analysis system (GE Healthcare, BI600, Boston, MA, USA).