Investigating PEDF and Inflammatory Pathways

Recombinant rat PEDF (GenBank accession number: NM_177927): Cusabio Biotech Co, Ltd (Wuhan, Hubei, China); The bacterial lipopolysaccharide (LPS) (L4391, O111:B4) and MPO colorimetric activity assay kits: Sigma (St. Louis, MO, USA). The LDH cytotoxicity assay kit and BCA protein concentration determination kit: Beyotime (Shanghai, China); Tissue and cell total protein extraction kits: Sangon Biotech (Shanghai, China). ELISA kits to measure TNF-α, IL-1β, and IL-6: Shanghai Renjie Biotechnology Co., Ltd. (Shanghai, China) [15 (link)]. ELISA kits to measure PEDF: Shanghai Yan Qi Biological Technology Co Ltd. The TUNEL kit, annexin V‑FITC/PI apoptosis detection kit, and DAPI staining solution: KeyGEN Biotech (Nanjing, Jiangsu, China). Anti-rabbit cleaved-caspase-3 (Catalog No. #9661), anti-rabbit p38 MAPK (Catalog No. #8690), anti-rabbit phospho-p38 MAPK (Catalog No. #4511), anti-rabbit NF-κB p65 (Catalog No. #8242), and anti-rabbit phospho-NF-κB p65 (Catalog No. #3033) antibodies: Cell Signaling Technology (Danvers, MA, USA). Anti-rabbit NLRP3 (Catalog No. ab263899), anti-rabbit PPAR-γ (Catalog No. ab178860) antibodies and PPAR-γ inhibitor (GW9662, Catalog No. ab141125): Abcam (Cambridge, MA, USA) [16 (link)]. Anti-rabbit RIP3 (Catalog No. 17563–1-AP) and anti-mouse β-tubulin (Catalog No. 66240–1-Ig) antibodies: Proteintech (Wuhan, Hubei, China).

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Xu L., Chen Y., Feng S., Liu Z., Ye Y., Zhou R, & Liu L. (2023). PEDF inhibits LPS-induced acute lung injury in rats and promotes lung epithelial cell survival by upregulating PPAR-γ. BMC Pulmonary Medicine, 23, 359.

Publication 2023

Recombinant rat PEDF ( MPO colorimetric activity assay kits LDH cytotoxicity assay kit BCA protein concentration determination kit TUNEL kit annexin V‑FITC/PI apoptosis detection kit DAPI staining solution

Annexin v fitc Antibodies Apoptosis Assay Bacterial Biological Caspase 3 Cell Colorimetric Cytotoxicity Dapi Elisa Gw9662 Il 1β Lipopolysaccharide Mouse Nf κb p65 P38 mapk Pedf Ppar γ Protein Rabbit Rip3 Tissue Tnf α Tubulin Tunel

Corresponding Organization :

Other organizations : Soochow University, Second Affiliated Hospital of Soochow University, Yangzhou University, Xuzhou Medical College, Second Hospital of Anhui Medical University, Anhui Medical University

Top 5 similar protocols

Variable analysis

independent variables

Recombinant rat PEDF (GenBank accession number: NM_177927)
Bacterial lipopolysaccharide (LPS) (L4391, O111:B4)

dependent variables

TNF-α, IL-1β, and IL-6 levels
PEDF levels
Apoptosis
Cleaved-caspase-3 levels
P38 MAPK and phospho-p38 MAPK levels
NF-κB p65 and phospho-NF-κB p65 levels
NLRP3 levels
PPAR-γ levels
RIP3 levels

control variables

MPO colorimetric activity
LDH cytotoxicity
Protein concentration

positive controls

Not specified

negative controls

Not specified

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