Journal: Kidney international
Article Title: Selective nanoparticle-mediated targeting of renal tubular Toll-like receptor 9 attenuates ischemic acute kidney injury.
doi: 10.1016/j.kint.2020.01.036
Figure Lengend Snippet: A. Representative images of terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) staining indicative of renal tubular apoptosis and counts of TUNEL positive kidney cells (B) in the kidneys of control MNP injected and MNPODN2088 injected mice subjected to sham-surgery (N=4) or to 30 min renal ischemia and 24 hr reperfusion (N=6, magnification 200X). Some mice were injected with MNP-ODN2088 6 hr before renal ischemia. A separate cohort of mice was injected with MNP-ODN2088 at the time of reperfusion or 1.5 hr after reperfusion. Another cohort of mice were injected with 5 mg/kg naked ODN2088 i.v. 6 hr before renal ischemia (N=5–6). C. We detected caspase 3 and caspase 8 activation by measuring cleaved caspase 3 and caspase 8 in the kidney lysates from sham-operated mice and mice subjected to renal IR injury after control MNP or MNP-ODN2088 treatment. *P<0.05 vs. control MNP injected mice subjected to sham surgery. #P<0.05 vs. control ODN mice subjected to renal IR. Error bars represent 1 SEM. D. We detected caspase 3 and caspase 8 activation in primary cultures of mouse renal proximal tubule cells treated with control ODN or with 5 mM ODN1668 (a selective mouse TLR9 activating ligand). Some proximal tubule cells were pretreated with MNPs encapsulating control ODN or MNPs encapsulating 1 mM ODN2088. *P<0.05 vs. control ODN treated mouse proximal tubule cells. #P<0.05 vs. ODN1668 treated mouse proximal tubule cells. Error bars represent 1 SEM.
Article Snippet: Confluent cells were treated with control oligonucleotides (ODN) or with selective 5 mM TLR9 agonist ligand ODN1668 (InvivoGen, CA) for 3 days as described 56 .
Techniques: End Labeling, TUNEL Assay, Staining, Injection, Activation Assay
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