Journal: CNS Neuroscience & Therapeutics
Article Title: The novel Nrf2 activator CDDO‐EA attenuates cerebral ischemic injury by promoting microglia/macrophage polarization toward M2 phenotype in mice
doi: 10.1111/cns.13496
Figure Lengend Snippet: CDDO‐EA increases expression of HO‐1, CD206, and CCL22, reduces CD16 and CD11b expression, and enhances microglial phagocytosis in LPS‐activated BV2 microglial cells. A, B, Cells were treated with CDDO‐EA (0, 50, 100, or 200 μg/mL), LPS (100 ng/mL), LPS (100 ng/mL) + CDDO‐EA (100 μg/mL), or LPS (100 ng/mL) + PPIX (10 μg/mL) for 24 h, lysates collected, and homogenates were immunoblotted with anti‐HO‐1 and anti‐β‐actin. C, D, BV2 cells were treated with LPS (100 ng/mL), LPS (100 ng/mL) + CDDO‐EA (100 μg/mL), or LPS + CDDO‐EA + PPIX (10 μg/mL) for 24 h, and the level of CD16, CD11b, CD206, and CCL22 mRNA were detected with real‐time quantitative PCR. E, BV2 cells were treated with LPS (100 ng/mL), LPS (100 ng/mL) + CDDO‐EA (100 μg/mL), or LPS + CDDO‐ EA + PPIX (10 μg/mL) for 21 h, fluorescent microspheres were added into the medium for 3 h, and then, the cells were stained with phalloidin to visualize F‐actin. The left panel is representative images of intra‐microglia fluorescence 3 h after fluorescent microspheres uptake. Scale bars = 50 μm (ns denotes not significant, * P < 0.05, ** P < 0.01, n = 3‐6)
Article Snippet: Cells were plated 24 hours prior to stimulation at a confluency of 80%, and treated with CDDO‐EA (0, 50, 100 and 200 μg/mL, HY‐12213, MedChemExpress), LPS (100 ng/mL, L4391, Sigma‐Aldrich), LPS (100 ng/mL) + CDDO‐EA (100 μg/mL), or LPS (100 ng/mL) + tin‐protoporphyrin IX (Sn‐PPIX, a specific HO‐1 inhibitor, 10 μg/mL, HY‐101194, MedChemExpress) for 24 hours.
Techniques: Expressing, Real-time Polymerase Chain Reaction, Staining, Fluorescence
