Journal: Arthritis Research & Therapy

Article Title: Sirt3 improves monosodium urate crystal-induced inflammation by suppressing Acod1 expression

doi: 10.1186/s13075-023-03107-6

Figure Lengend Snippet: MSU crystals stimulation reduced the expression and deacetylation activity of Sirt3. a Sirt3 mRNA and protein levels were detected in PBMCs of health controls (HC), patients with acute gout (AG), and intermittent gout (IG). Sirt3 mRNA detection (HC, n = 24), (AG, n = 24), (IG, n = 24); GAPDH was used as an internal reference. Sirt3 protein detection (HC, n = 6), (AG, n = 6), (IG, n = 6), TUBULIN as an internal reference. b BMDMs were respectively stimulated with C16:0, MSU, or C16:0 + MSU for 12 h and Western blot was used to detect Sirt3 protein expression. c The protein levels of Sirt3 in the mouse paw treated with MSU crystals or vehicle. Data presented as mean ± SD ( n = 5 mice per group). * P < 0.05. NS means no statistical difference. d Sirt3 agonists (Viniferin) inhibited the acetylation level of mitochondrial proteins in BMDMs treated with Viniferin (1 μM) and C16:0 + MSU crystals for 12 h. Acetyl-conjugated proteins in mitochondrial proteins were blotted with acetylated-lysine antibody. e Viniferin treatment decreased K68 lysine acetylation of mitochondrial SOD2 protein and mitochondrial proteins were analyzed by Western blot in BMDMs. f The activity of SOD in the mitochondrial proteins of BMDMs was detected with WST-8 method. Data are mean ± SD. Three independent repetitive experiments were conducted for each result. *P < 0.05. NS represents P > 0.05

Article Snippet: HY-133987, Mito-TEMPO, and Viniferin were purchased from MedChemExpress (MCE).

Techniques: Expressing, Activity Assay, Western Blot

Journal: Marine Drugs

Article Title: Fucoxanthin Enhances Cisplatin-Induced Cytotoxicity via NFκB-Mediated Pathway and Downregulates DNA Repair Gene Expression in Human Hepatoma HepG2 Cells

doi: 10.3390/md11010050

Figure Lengend Snippet: ( A ) The ratio of Bax/Bcl-2 mRNA in HepG2 cells pretreated with fucoxanthin (1–10 μM) for 24 h followed by incubation with cisplatin (10 μM) for 24 h. ( B ) The ratio of Bax/Bcl-2 mRNA in HepG2 cells pretreated with fucoxanthin (5 μM) for 24 h followed by incubation with NFκB activation inhibitor (NAI, 20 μΜ) for 2 h and then treated cisplatin (10 μM) for 24 h. Values are means ± SD, n = 3; means without a common letter differ significantly, P < 0.05.

Article Snippet: NFκB activation inhibitor was purchased from Merck Millipore (Billerica, MA, USA).

Techniques: Incubation, Activation Assay

Journal: Molecular Therapy

Article Title: Local Delivery of miR-21 Stabilizes Fibrous Caps in Vulnerable Atherosclerotic Lesions

doi: 10.1016/j.ymthe.2018.01.011

Figure Lengend Snippet: miR-21 Deficiency Activates NF-kB Nuclear Translocation and Enhances OxLDL Uptake in Mouse Peritoneal Macrophages

Article Snippet: To investigate the mechanism, 9 μM NF-kB nucleus translocation inhibitor SN50 (Merck) was added to cultured macrophages before oxLDL loading.

Techniques: Translocation Assay

Journal: Nature Communications

Article Title: Dietary cholesterol directly induces acute inflammasome-dependent intestinal inflammation

doi: 10.1038/ncomms6864

Figure Lengend Snippet: ( a ) Schematic representation of the two signal model of activation of the inflammasome. Magenta boxes indicate chemical or genetic tools (MO) used and turquoise diamonds indicate reporter activity tools. ( b ) Total number of intestinal L-Plastin+ cells in ezetimibe (25 μM) or DMSO-treated larvae. n ≥30, one representative experiment of three. One-way analysis of variance (ANOVA). ( c ) Total number of intestinal L-plastin+ cells in NAI inhibitor (150 nM)-treated larvae. n ≥16, one representative experiments of two. One-way ANOVA. ( d ) Total number of intestinal L-plastin+ cells in larvae reared in conventional (CONV) or GF conditions and fed sterile HCD, ZM control diet or unfed. n ≥8, pooled from two experiments. One-way ANOVA. ( e ) Total number of intestinal L-plastin+ cells in NADPH oxidase inhibitor VAS-28701 (1 μM)-treated larvae. n ≥28, pooled from two experiments. Kruskal–Wallis test. ( f ) Total number of intestinal L-plastin+ cells in Cathepsin B inhibitor Ca-074-Me (100 μM)-treated larvae. n ≥34, pooled from two experiments. Kruskal–Wallis test. ( g ) Total number of intestinal L-Plastin+ cells in Caspase-1 inhibitor N -AcetylWEHD-al-treated larvae. n ≥29, pooled from two experiments. Kruskal–Wallis test. From b to g , each dot represents one individual larva. Error bars are s.e.m. *** P <0.001 and ** P <0.01.

Article Snippet: WT zebrafish larvae (5 dpf) were pretreated in 25 μM ezetimibe (Sequoia Research Ltd) overnight, followed by feeding the next day for 6 h. For all other inhibitors, WT larvae (6 dpf) were pretreated for 30 min in Caspase-1 and 5 inhibitor N -Acetyl WEHD-al (Sigma), Cathepsin B inhibitor Ca-074-Me (Calbiochem), NADPH oxidase inhibitor VAS-2870 (Enzo Life Sciences) and NF-κB-activation inhibitor NAI (Merck), followed by feeding for 6 h. The inhibitors were administered into the water and were present for the duration of the feeding.

Techniques: Activation Assay, Activity Assay