Journal: International Journal of Molecular Sciences
Article Title: Oxyresveratrol Inhibits IL-1β-Induced Inflammation via Suppressing AKT and ERK1/2 Activation in Human Microglia, HMC3
Figure Lengend Snippet: The effects of OXY on regulating AKT phosphorylation in cells stimulated with IL-1β. Representative images from immunofluorescence study showing AKT phosphorylation at Ser473 (green) of untreated cells ( a–c ), IL-1β-stimulated cells ( d–f ), IL-1β-stimulated cells with the presence of OXY at 40 μM ( g–i ), or DMSO ( j–l ). Nuclei (blue) were stained with DAPI. The micrographs were captured at 100× magnification, and data are representatives of three replicates.
Article Snippet: After blocked with 5% skim milk in a mixture of tris-buffered saline and Tween 20 (TBS-T) (0.02 M Tris-HCl, pH 7.6, 0.0137 M NaCl, and 0.05% Tween 20) (all reagents from Sigma-Aldrich, Saint Louis, MO, USA) at RT for 1 h, membranes were incubated with an appropriate primary antibody (Cell Signaling Technology, Boston, MA, USA) at 4 °C for 24 h. Primary antibodies used in this study included an anti-NF-κB p65 (D14E12) antibody (catalog number 8242), a mouse anti-IκBα (L35A5) antibody (catalog number 4814), a phosphospecific rabbit anti-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (catalog number 4370), a mouse anti-p44/42 MAPK (Erk1/2) antibody (catalog number 9107), a phosphospecific mouse anti-p38 MAPK (Thr180/Tyr182) antibody (catalog number 9216), a rabbit anti-p38 MAPK (D13E1) antibody (catalog number 8690), a phosphospecific rabbit anti-SAPK/JNK (Thr183/Tyr185) antibody (catalog number 4668), a rabbit anti-SAPK/JNK antibody (catalog number 9252), a phosphospecific rabbit anti-AKT (Ser473) antibody (catalog number 4060), a mouse anti-AKT (40D4) antibody (catalog number 2920), a phosphospecific rabbit p70 S6 kinase (Thr389) antibody (catalog number 97596), or a mouse anti-β-actin antibody (catalog number MA1115) (Boster Biological Technology, Pleasanton, CA, USA).
Techniques: Immunofluorescence, Staining