Figure 3: 25-HC induces microglial recruitment and IL-1β production.

(a) Illustration of stereotaxic injection of 25-HC (100 μM) or vehicle (0.1% ethanol in PBS) into the corpus callosum of mouse brain. (b) Representative immunofluorescence image of a coronal brain section stained with anti-Iba-1 antibody (green). DAPI represents the nuclear signal (blue). Scale bar, 200 μm. (c) Quantification of Iba-1-positive cells in the boxed areas of vehicle- or 25-HC-injected regions in b. (n=8) (d) Quantification of IL-1β in the brain homogenates of vehicle- or 25-HC-injected hemisphere. (n=11) (e) Representative immunofluorescence image of brain section co-stained with FLICA (green) and anti-Iba-1 antibody (red) after injection of vehicle or 25-HC. Magnified immunofluorescence images of the boxed areas in the upper panel are displayed in the lower panel: 25-HC-injected area (right) or vehicle-injected area (left). Scale bars, 200 μm (upper); or 50 μm (lower). (f,g) Quantification of IL-1β or IL-6 in the culture supernatants of mouse BMDMs untreated or treated with 25-HC (70 μM) or VLCFA (70 μM) for 10 h in the presence of LPS (0.25 μg ml⁻¹, last 4 h) as indicated. (n=6). For all panels, **P<0.01 and ***P<0.001.