Journal: Molecules

Article Title: Steppogenin Isolated from Cudrania tricuspidata Shows Antineuroinflammatory Effects via NF-κB and MAPK Pathways in LPS-Stimulated BV2 and Primary Rat Microglial Cells

doi: 10.3390/molecules22122130

Figure Lengend Snippet: Figure 9. The effects of steppogenin (1) on IκB-α phosphorylation and degradation (A), NF-κB activation (B,C), and NF-κB localization (D) in LPS-stimulated primary rat microglial cells. (A–D) The cells were pretreated for 3 h with the indicated concentrations of 1 and then stimulated for 1 h with LPS (1 µg/mL). Total proteins were prepared and the western blot analysis was performed using specific IκB-α, p-IκB-α p65, and p50 antibodies. A commercially available NF-κB ELISA (Active Motif) was used to test the nuclear extracts and determine the degree of NF-κB binding. The data are presented as the mean ± SD of three experiments. The band intensities were quantified by densitometry and normalized to the intensity of β-actin or PCNA; the normalized values are presented below each band.

Article Snippet: An aliquot of the supernatant (100 μL) was mixed with an equal volume of Griess reagent (Solution A: 222488, Solution B: S438081; Sigma-Aldrich) and the absorbance of the mixture at 525 nm was determined by using an enzyme-linked immunosorbent assay (ELISA) plate reader (Bio-Rad model 680, Hercules, CA, USA) [37].

Techniques: Phospho-proteomics, Activation Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Binding Assay