Journal: PLoS ONE

Article Title: Stearoyl-CoA Desaturase-1 (SCD1) Augments Saturated Fatty Acid-Induced Lipid Accumulation and Inhibits Apoptosis in Cardiac Myocytes

doi: 10.1371/journal.pone.0033283

Figure Lengend Snippet: (A): FA oxidation was significantly increased in rat neonatal cardiac myocytes treated with 100 µM palmitic acid (PA). (B): On the other hand, glucose oxidation was significantly reduced in cardiac myocytes treated with PA (100 µM). (C): PA (50 to 100 µM) induced excessive FA oxidation was attenuated in cardiac myocytes transduced with a MOI of 20 of Ad-SCD1 (▪) in comparison with control cells transduced with Ad-LacZ (□). (D): PA (50 to 100 µM) -induced suppression of glucose oxidation recovered in cardiac myocytes transduced with Ad-SCD1 (▪) in comparison with control cells transduced with Ad-LacZ (□). BSA was used as a vehicle. Each sample was counted in a scintillation counter (cpm) and data are shown as the mean ± SD. **P<0.01 vs. Ad-LacZ in each PA concentration. (E); Phosphorylation levels of AMPK and ACC decreased in cardiac myocytes transduced with an MOI of 20 of Ad-SCD1 in comparison with control cells transduced with Ad-LacZ. Oligomycin (1 µM) was used as an AMPK and ACC activator.

Article Snippet: The mixture was rotated at 4°C for 15 min and centrifuged at 14000 rpm for 15 min. Western blot analysis was performed according to standard procedures using the following primary antibodies: rabbit monoclonal phosphor-AMPK, AMPK, phosphor-ACC, ACC, cleaved-caspase 3, caspase 3 mAb (Cell Signaling) and β-actin (Santa cruz biotechnology, CA, USA).

Techniques: Transduction, Concentration Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Nicotinamide Mononucleotide Ameliorates Cellular Senescence and Inflammation Caused by Sodium Iodate in RPE

doi: 10.1155/2022/5961123

Figure Lengend Snippet: (a–d) ARPE-19 cells were treated with 1 and 2.5 mM NaIO 3 for 48 h. (a) Mitochondrial membrane potential (MMP) in ARPE-19 cells was demonstrated by JC-1 staining, and the results were obtained by fluorescent microscopy. The red-to-green fluorescence intensity was used to quantitatively evaluate the mitochondrial potential; scale bar = 50 μ m ( n = 3). (b) Cellular ATP levels in ARPE-19 cells ( n = 5). (c) NAD + levels in ARPE-19 cells ( n = 3). (d) Expression of AMPK α and p-AMPK α proteins in ARPE-19 cells. (e) Immunofluorescence assay of mitochondrial outer membrane marker Tom20 was performed by confocal microscope. Cells were treated with 1 mM NaIO 3 for 48 hr; scale bar = 10 μ m ( n = 3) The mitochondrial branches length were measured by ImageJ. Data represent mean ± SD of at least three independent experiments, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, compared versus control.

Article Snippet: The antibodies specific for p21 (#37543), Sirt1 (#2493), AMPK α (#5831), phosphor-AMPK α (Thr172, #2535), total-Akt (#4685), phosphor-Akt (Ser473, #4060), mTOR (#2983), and phosphor-mTOR (Ser2448, #5536), λ -H2AX (#2577) was purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Staining, Microscopy, Fluorescence, Expressing, Immunofluorescence, Marker