Journal: Journal of the American Society of Nephrology : JASN

Article Title: Drugs Repurposed as Antiferroptosis Agents Suppress Organ Damage, Including AKI, by Functioning as Lipid Peroxyl Radical Scavengers

doi: 10.1681/ASN.2019060570

Figure Lengend Snippet: The antiferroptotic effects of the drugs on cells derived from various organs and species. (A) Antiferroptotic effect of the drugs on various cell lines: NRK49F (100 μM BSO, 96 hours), HK-2 (5 μM erastin, 48 hours), HUPEC (0.5 μM erastin, 48 hours), C2C12 (200 μM BSO, 60 hours). Panc-1 (5 μM erastin, 30 hours), and MDA-MB-231 (10 μM erastin, 48 hours). n=3. (B) Prevention of glutamate-induced neuronal cytotoxicity in HT22 cells by drugs. Cells were exposed to 4 mM glutamate for 24 hours. n=3. (C) Antiferroptotic effects of phenothiazine-derived drugs and rifamycin derivatives evaluated in BSO (100 μM)-treated H9C2 cells. n=3. P values were determined by one-way ANOVA, Dunnett’s. *P<0.05. Data are mean±SEM.

Article Snippet: H9C2 (rat cardiomyoblast), NRK49F (rat kidney fibroblast), HK2 (human kidney tubular cell), C2C12 (mouse myoblast), MDA-MB-231 (human breast adenocarcinoma), NRK52E (rat kidney tubular cell), and LLC-PK1 (porcine kidney tubular cell) cells were obtained from ATCC.

Techniques: Derivative Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Class I and III phosphatidylinositol 3'-kinase play distinct roles in TLR signaling pathway.

doi: 10.4049/jimmunol.176.10.5943

Figure Lengend Snippet: FIGURE 1. PI3K inhibitor wortmannin inhibits CpG ODN-mediated responses in a dose-dependent manner. A, mTLR9–293T or RAW264.7 cells were transfected with p5xNF-B luciferase. After overnight transfec- tion, cells were stimulated with CpG ODN (1.5 M) for 8 h in the presence of various concentrations of wortmannin, and their NF-B luciferase ac- tivities were then measured. B, RAW264.7 cells were pretreated with var- ious concentrations of wortmannin for 1 h, followed by CpG ODN (1.5 M) stimulation for 20 h. NO, IL-6, IL-12, and TNF- levels in culture supernatants were measured by ELISA. Data represent the mean SD of three experiments. , p 0.005; , p 0.001.

Article Snippet: Cell culture and cell treatment The 293T human embryonic kidney fibroblasts and mouse RAW264.7 macrophages were obtained from the American Type Culture Collection.

Techniques: Transfection, Luciferase, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Class I and III phosphatidylinositol 3'-kinase play distinct roles in TLR signaling pathway.

doi: 10.4049/jimmunol.176.10.5943

Figure Lengend Snippet: FIGURE 6. Class I PI3K kinase-dead mutant impaired NF-B activa- tion induced by wild-type MyD88 overexpression. In addition to the NF- B-driven luciferase gene, wild-type MyD88-overexpressing 293T cells were transfected with kinase-deficient plasmid hVSP34kd or M-p110- kin-myc. After 24-h transfection, NF-B luciferase activities were mea- sured. Data represent the mean SD of three experiments.

Article Snippet: Cell culture and cell treatment The 293T human embryonic kidney fibroblasts and mouse RAW264.7 macrophages were obtained from the American Type Culture Collection.

Techniques: Mutagenesis, Over Expression, Luciferase, Transfection, Plasmid Preparation