sirtuin 3  (Cell Signaling Technology Inc)

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Resolvin D4 mitigates lipopolysaccharide-induced lung injury in mice.

doi: 10.1016/j.plefa.2024.102652

Figure Lengend Snippet: Fig. 3. The effect of RvD4 administration on the cell apoptosis of alveolar epithelial cells and the protein expression of Zo-1 and Sirt3 in the lungs of LPS-injected mice. A: Representative profiles of TUNEL staining (red) of lung sections 24 h after LPS injection (5 mg/kg). Note that 4′,6-diamidino-2-phenylindole (DAPI) was used in the counterstaining of the nucleus (blue). Scale bars: 50 μm. Three high-power fields per slide were randomly selected, and the number of TUNEL-positive cells and DAPI-positive cells were counted. The percentages of TUNEL-positive cells were obtained by dividing the number of TUNEL-positive cells by the number of DAPI- positive cells. TUNEL-positive cells (arrows) were visualized in lung tissues of LPS-injected mice. Data are means ± SEM of 4 mice per group. *p < 0.05. B,C: Immunoblots of Zonula occludens-1 (Zo-1) and sirtuin-3 (Sirt3) from the PBS control, LPS/saline, and LPS/RvD4 groups. The quantitative comparisons were per formed using β-actin as the loading control (PBS control, n = 7; LPS/saline, n = 6; LPS/RvD4, n = 8). Values are mean ± SEM, analyzed by one-way ANOVA and then Dunnett multiple comparison test. *p < 0.05 vs. the LPS/saline group.

Article Snippet: We measured the expression levels of proteins in the lysates of lung tissue by western blotting, using antibodies recognizing the following proteins: β-actin (Sigma-Aldrich Japan), Zonula occludens-1 (Zo-1) (Cell Signaling Technology Japan, Tokyo), and sirtuin-3 (Sirt3) (Cell Signaling Technology Japan).

Techniques: Expressing, Injection, TUNEL Assay, Staining, Western Blot, Control, Saline, Comparison

Journal: Prostaglandins, leukotrienes, and essential fatty acids

Article Title: Resolvin D4 mitigates lipopolysaccharide-induced lung injury in mice.

doi: 10.1016/j.plefa.2024.102652

Figure Lengend Snippet: Fig. 5. The silencing of Sirtuin-3 (SIRT3) abolished the anti-inflammatory effect and retention of cell integrity in RvD4-treated BEAS-2B cells. A: The silencing efficiency of SIRT3 was determined by quantitative real-time PCR. Data are shown as mean ± SEM (n= 4 per group). *p < 0.05. B: The mRNA expression levels of IL- 1β and IL-6 in BEAS-2B cells transfected with SIRT3 siRNA (si-SIRT3) or scrambled siRNA (si-scr) at 3 h after LPS stimulation are shown. Gene expressions were normalized with hypoxanthine phosphoribosyltransferase 1 (HPRT1). Data are shown as mean ± SEM (n= 3 per group). *p < 0.05. n.s.: not significant. C: Time- dependent changes in the TER from baseline in BEAS-2B cells exposed to PBS or LPS. Data are shown as mean ± SEM, analyzed by one-way ANOVA and then Dunnett multiple comparison test (n= 4 per group). *p < 0.05. n.s.: not significant.

Article Snippet: We measured the expression levels of proteins in the lysates of lung tissue by western blotting, using antibodies recognizing the following proteins: β-actin (Sigma-Aldrich Japan), Zonula occludens-1 (Zo-1) (Cell Signaling Technology Japan, Tokyo), and sirtuin-3 (Sirt3) (Cell Signaling Technology Japan).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Transfection, Comparison

Journal: Life

Article Title: Supplementation of Nicotinic Acid and Its Derivatives Up-Regulates Cellular NAD + Level Rather than Nicotinamide Derivatives in Cultured Normal Human Epidermal Keratinocytes

doi: 10.3390/life14030413

Figure Lengend Snippet: Ablation of mitochondrial ROS by the supplementation of nicotinic acid ( A ) Representative images of the effects of NA on the mitochondrial markers superoxide dismutase (SOD) 2 and sirtuin 3 (SIRT3). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is shown as the loading control. ( B , C ) Quantification of SOD2 ( B ) and SIRT3 ( C ) immunoblot bands on ( A ). The data are divided by the intensity of GAPDH. Individual quantified values are represented as gray dots on the bar chart. ( D ) Representative images of the effect of NA on mitochondrial ROS recovery activity. Mitochondria and mitochondrial ROS were stained by green and red fluorescent dye, and 50 μM mito-TEMPO was used as a positive control. R: 1 μM rotenone. The red/green pixel ratio (R/G ratio) is depicted by a histogram, and the means of the histograms are shown in ( E ). All data are expressed as the mean ± SE of at least three independent experiments. The statistical analyses were performed by a one-way ANOVA followed by Dunnett’s test. * p < 0.05.

Article Snippet: The superoxide dismutase 2 (SOD2) and sirtuin 3 (SIRT3) antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Control, Western Blot, Activity Assay, Staining, Positive Control