Journal: Molecular Medicine Reports

Article Title: MicroRNA-217 is involved in the progression of atherosclerosis through regulating inflammatory responses by targeting sirtuin 1

doi: 10.3892/mmr.2019.10581

Figure Lengend Snippet: Effects of miR-217 inhibitor on SIRT1/AMPK-α/NF-κB pathway in THP-1 macrophages. THP-1 macrophages were pre-transfected with miR-217 inhibitor, inhibitor control, or miR-217 inhibitor + SIRT1-siRNA and treated with ox-LDL. (A) The protein levels of SIRT1, p-AMPK-α, AMPK-α, p-p65, and p65 in THP-1 macrophages was determined using western blotting. (B) the mRNA level of SIRT1 in THP-1 macrophages was determined using reverse transcription-quantitative PCR. The ratio of (C) p-AMPK-α/AMPK-α and (D) p-p65/p65 was calculated. **P<0.01 vs. Control; ## P<0.01 vs. ox-LDL; && P<0.01 vs. Inhibitor. AMPK-α, 5′-AMP-activated kinase α; miR-217, microRNA-217; ox-LDL, oxidized low density lipoprotein; p, phosphorylated; SIRT1, sirtuin 1.

Article Snippet: Following blocking in 5% skimmed milk at room temperature for 1.5 h, the membranes were incubated with primary antibodies against SIRT1 (120 kDa; 1:1,000; cat. no. 9475; Cell Signaling Technology, Inc.), phosphorylated (p)-AMP-activated protein kinase α (AMPK-α; 62 kDa; 1:1,000; cat. no. 50081; Cell Signaling Technology, Inc.), AMPK-α (62 kDa; 1:1,000; cat. no. 5831; Cell Signaling Technology, Inc.), p-NF-κB p65 (65 kDa; 1:1,000; cat. no. 3033; Cell Signaling Technology, Inc.), p65 (65 kDa; 1:1,000; cat. no. 8242; Cell Signaling Technology, Inc.) and β-actin (45 kDa; 1:1,000; cat. no. 4970; Cell Signaling Technology, Inc.) overnight at 4°C.

Techniques: Transfection, Control, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Journal of Korean Medical Science

Article Title: Pregabalin as a Neuroprotector after Spinal Cord Injury in Rats: Biochemical Analysis and Effect on Glial Cells

doi: 10.3346/jkms.2011.26.3.404

Figure Lengend Snippet: The effect of intraperitoneal injection of pregabalin (30 mg/kg) on the expression of phosphorylated p38 MAPK in contusive spinal cord injury. The values were examined on post-injury day 7. The methylprednisolone and pregabalin treatment groups demonstrate a reduced expression of phosphorylated p38 MAPK as compared to that of the control group. However, ANOVA indicated no statistically significant difference (f = 0.65, P = 0.58, n = 10 for the experimental group, n = 3 for the sham group). The following Bonferroni post hoc analysis also indicated no statistical difference between the groups.

Article Snippet: The antibodies used for immunoblotting were as follows: anti-rabbit activated caspase-3 antibody (1:1,000; Cell Signaling Technology, Danvers, MA, USA), anti-rabbit phosphorylated p-38 MAPK (1:1,000; Cell Signaling Technology), anti-rabbit p-38 MAPK (1: 1,000; Cell Signaling Technology) and anti-mouse Bcl-2 (1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Injection, Expressing, Control

Journal: Molecular Medicine Reports

Article Title: Inhibition of mTOR signaling protects photoreceptor cells against serum deprivation by reducing oxidative stress and inducing G 2 /M cell cycle arrest

doi: 10.3892/mmr.2016.5011

Figure Lengend Snippet: Inhibition of mammalian target of rapamycin signaling attenuates serum deprivation-induced cell death. (A–C) Cell death was evaluated using propidium iodide (PI) staining (scale bars = 100 µ m). (A) 661W control cells were cultured under normal conditions. (B) 661W cells were cultured in serum-free medium for 3 days. (C) 661W cells were treated with 100 nM rapamycin alongside serum deprivation for 3 days. Scale bar, 100 µ m. (D) Quantification of cell death via PI staining. *** P<0.001 vs. the serum-deprived group. Assays were performed in quadruplicate, and data are presented as the mean ± standard error of the mean. Normal, cells cultured in normal medium; SD, cells cultured in serum-free medium; SD+Rap, cells cultured in serum-free medium and treated with 100 nM rapamycin. (E) Treatment with rapamycin (100 nM) reduced phosphorylated (p)-P70S6 kinase (P70S6K) and p-eukaryotic initiation factor 4E-binding protein (4EBP1) expression on days 2 and 4 of serum deprivation.

Article Snippet: Rabbit antibodies against phosphorylated (p)-P70S6 kinase (P70S6K) (cat. no. 11284), p-4EBP1 (cat. no. 11223) and mouse β-actin (cat. no. 21800-1) were purchased from Signalway Antibody LLC (College Park, MD, USA).

Techniques: Inhibition, Staining, Control, Cell Culture, Binding Assay, Expressing