Journal: Diabetes

Article Title: Loss of NADPH Oxidase–Derived Superoxide Skews Macrophage Phenotypes to Delay Type 1 Diabetes

doi: 10.2337/db14-0929

Figure Lengend Snippet: NOD.Ncf1m1J islets exhibit lowered M1 and elevated M2 levels of macrophage markers. Purified islet whole-cell lysates from prediabetic 16-week-old NOD and NOD.Ncf1m1J mice were used in an immunoblot analysis for P-STAT6 (Y641) and STAT6 (A), iNOS (B), and Arg-1 (C). Densitometry statistics for Western blotting represent the average of three experiments. ***P < 0.001; *P < 0.05. WT, wild type.

Article Snippet: For Western blotting, whole-cell lysates were probed with antibodies against iNOS (Novus Biologicals), Arg-1, STAT6 (Cell Signaling), P-STAT6 (Y641) (BD Bioscience), or β-actin (Sigma-Aldrich) as described ( 18 ).

Techniques: Purification, Western Blot

Journal: British Journal of Nutrition

Article Title: Treatment with oligonol, a low-molecular polyphenol derived from lychee fruit, attenuates diabetes-induced hepatic damage through regulation of oxidative stress and lipid metabolism

doi: 10.1017/s0007114511001322

Figure Lengend Snippet: Fig. 2. NF-kBp65 (A), cyclo-oxygenase-2 (COX-2) (B) and inducible NO synthase (iNOS) (C) expressions in the liver. m/m, Misty; db/db, diabetic; Veh, db/db vehicle-treated mice; Oligo-10, db/db mice treated with oligonol at 10 mg/kg body weight; Oligo-20, db/db mice treated with oligonol at 20 mg/kg body weight. Values are means (n 6 or n 10), with standard errors represented by vertical bars. a,b Mean values with unlike letters were significantly different (P,0·05; Duncan’s test).

Article Snippet: Rabbit polyclonal antibodies against PPARa, SREBP-1, SREBP-2, NF-kBp65 and RAGE, and mouse monoclonal antibody against cyclo-oxygenase-2 (COX-2) and inducible NO synthase (iNOS) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques:

Journal: British Journal of Nutrition

Article Title: Treatment with oligonol, a low-molecular polyphenol derived from lychee fruit, attenuates diabetes-induced hepatic damage through regulation of oxidative stress and lipid metabolism

doi: 10.1017/s0007114511001322

Figure Lengend Snippet: Fig. 5. Hepatic mRNA expressions of acetyl-CoA carboxylase (ACC) (A), fatty acid synthase (FAS) (B) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) (C). m/m, Misty; db/db, diabetic; Veh, db/db vehicle-treated mice; Oligo-10, db/db mice treated with oligonol at 10 mg/kg body weight; Oligo-20, db/db mice treated with oligonol at 20 mg/kg body weight. Values are means (n 6 or n 10), with standard errors represented by vertical bars. a,b Mean values with unlike letters were significantly different (P,0·05; Duncan’s test).

Article Snippet: Rabbit polyclonal antibodies against PPARa, SREBP-1, SREBP-2, NF-kBp65 and RAGE, and mouse monoclonal antibody against cyclo-oxygenase-2 (COX-2) and inducible NO synthase (iNOS) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques:

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: S100A9 -/- alleviates LPS-induced acute lung injury by regulating M1 macrophage polarization and inhibiting pyroptosis via the TLR4/MyD88/NFκB signaling axis.

doi: 10.1016/j.biopha.2024.116233

Figure Lengend Snippet: Fig. 7. S100A9-gene deficiency down-regulated M1 polarization and TLR4/MyD88/NFκB signaling axis expression in BMDMs. (A-B) Immunofluorescence staining analysis the expression of iNOS (N = 5) and TLR4/NFκB (N = 3) in BMDMs respectively from the WTCTR, A9-/-CTR, WTLPS, and A9-/-LPS. All scale bar, 50 µm. (C-E) Comparison of the percentage of iNOS-/TLR4-/NFκB-positive cells. (F-J) Western blot analysis of the expression of iNOS/TLR4/MyD88/NFκB to β-actin in BMDMs from the WTCTR, A9-/-CTR, WTLPS, and A9-/-LPS (N = 3). WTCTR: BMDMs derived from WT mice cultured in 1640 medium; A9-/-CTR: BMDM cells derived from S100A9 KO mice cultured in 1640 medium; WTLPS: BMDMs derived from WT mice cultured in 1640 medium containing LPS (1 μg/ml); A9-/-LPS: BMDMs derived from S100A9 KO mice cultured in 1640 medium containing LPS (1 μg/ml). ns, no significance, * P < 0.05, * * P < 0.01, * ** P < 0.001, one-way ANOVA (Bonferroni post-hoc tests for subgroup comparisons).

Article Snippet: Overnight incubation at 4 ◦C with primary antibodies against NLRP3 (1:100, Abcam, ab263899, UK), caspase1 (1:100, C. Gong et al. Biomedicine & Pharmacotherapy 172 (2024) 116233 Proteintech, cat# 22915–1-AP, China), TLR4 (1:100, Abcam, ab22048, UK), Phospho-NFκB (1:100, CST, 3033, USA), and iNOS (1:200, CST, 13120, USA) was followed by a 1 h incubation with appropriate secondary antibodies (Abcam, ab150113 USA; Abcam, ab150080 USA).

Techniques: Expressing, Immunofluorescence, Staining, Comparison, Western Blot, Derivative Assay, Cell Culture

Journal: Journal of Biomedicine and Biotechnology

Article Title: Regulation of P-Glycoprotein in Renal Proximal Tubule Epithelial Cells by LPS and TNF- α

doi: 10.1155/2010/525180

Figure Lengend Snippet: Expression of iNOS and effect of NO on Abcb1/P-gp activity in GERP cells. ((a) n = 4) NOSII expression in GERP cells after exposure to either 10 μ g/mL LPS (closed bars), 10 ng/mL TNF- α (striped bars) or both 10 μ g/mL LPS and 10 ng/mL TNF- α (chequered bars) for 2, 6 or 24 hours ( n = 4). mRNA levels were determined with RQ-PCR and expression was normalized for the GAPDH C T value. ((b) n = 4) iNOS protein expression in GERP cells (control; lanes 1 and 2) after exposure to either 10 μ g/mL LPS (lanes 3 and 4), 10 ng/mL TNF- α (lanes 5 and 6) or both 10 μ g/mL LPS and 10 ng/mL TNF- α (lanes 7 and 8) for 6 hours. ((c) n = 3–5) P-gp transport activity in GERP cells after exposure to 10 ng/mL TNF- α or both 10 μ g/mL LPS and 10 ng/mL TNF- α in combination with 50 or 10 μ M of the NO-scavenger PTIO, respectively, for 24 hours compared to control (cells exposed to culture medium). ((d) n = 4) P-gp protein expression in GERP cells (control; lanes 1 and 2) after exposure to 100 μ M SNP (lanes 3 and 4) for 1 hour with 5 hours recovery. ((e) n = 4) P-gp transport activity in GERP cells, compared control, after exposure to 100 or 500 μ M SNP for 6 hours or for 1 hour with a subsequent recovery period of 5 hours. Total cell fractions of GERP cells were used and expression of iNOS (b) or P-gp (d) was determined by Western blotting. Relative pixel intensities (ratio iNOS/ β -actin (b) and ratio P-gp/ β -actin (d)) were determined through image analysis. Data are expressed as mean ± SEM. Significantly different compared to control (*: P < .05, **; P < .01, ***: P < .001).

Article Snippet: Different rat genes ( GAPDH : (Rn99999916_S1), Abcb1b : (Rn00561753_m1), NOSII : (Rn00561646_m1), or Ednrb/endothelin receptor B : Rn00569139_m1) were amplified with a pre-developed Gene Expression Assay, provided by Applied Biosystems.

Techniques: Expressing, Activity Assay, Control, Western Blot