Journal: eLife

Article Title: Unsupervised machine learning reveals risk stratifying glioblastoma tumor cells

doi: 10.7554/eLife.56879

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-p-AKT (S473) (rabbit-monoclonal) , Fluidigm , RRID: AB_2811246 Cat#3152005A Clone: D9E , MC (1:100).

Techniques: Isolation, Software

Journal: Molecules

Article Title: Anti-Metabolic Syndrome Effects of Fucoidan from Fucus vesiculosus via Reactive Oxygen Species-Mediated Regulation of JNK, Akt, and AMPK Signaling

doi: 10.3390/molecules24183319

Figure Lengend Snippet: Effects of fucoidan from Fucus vesiculosus (FvF) on relieving insulin resistance (IR) in HepG2 cells. Effects of sodium palmitate (PA) on cellular glucose consumption ( a ). Cells were treated with a concentration range of PA for 24 h. Effects of FvF on glucose consumption in IR cells ( b ). Cells were treated with Metf (2 mM) or FvF (100 μg/mL) in the presence of 100 μM PA for 24 h. ( c ). Reactive oxygen species (ROS) was detected by in situ dihydroethidium (DHE) staining (200×). C, control group; M, cells were treated with 100 μM PA for 24 h; Metf and FvF, cells were treated with metformin (2 mM) or FvF (100 μg/mL) in the presence of 100 μM PA for 24 h. Phosphorylation of c-Jun N-terminal kinase (pJNK) ( d ) and phosphorylation of protein kinase B (pAkt) ( e ) protein levels changed between different treatment groups. C, control group; M, cells treated with 100 μM PA for 24 h; Metf and FvF, cells treated with 100 μM PA for 24 h then incubated with metformin (2 mM) or FvF (100 μg/mL) for another 6 h. Data are expressed as the mean ± SEM. Differences were assessed by ANOVAs and statistical results are denoted as follows: * p < 0.05 versus the control group; # p < 0.05 versus the model group.

Article Snippet: After that, HepG2 cells were rinsed with PBS and lysed in ice-cold RIPA buffer containing protease inhibitor and phosphatase inhibitors (Roche) for 30 min. After denaturation with 5× loading buffer at 100 °C for 10 min, proteins were electrophoresed on 10% SDS-PAGE and transferred to a PVDF membrane (0.22 μm) that was subsequently blocked with 5% ( w / v ) non-fat milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 2 h. The membranes were incubated with primary antibodies directed against JNK, pJNK (T183/T185), Akt, pAkt (S473), AMPK, pAMPK (T172), HMGCR, pHMGCR (S872) (Merck Millipore, Billerica, MA, USA), SREBP-1C (Abcam, HongKang, China), ACC, pACC (S79), and β-actin antibodies in 5% ( w / v ) BSA in TBST overnight at 4 °C.

Techniques: Concentration Assay, In Situ, Staining, Control, Phospho-proteomics, Incubation