Figure 6: 25-HC promotes the microglial recruitment and oligodendrocyte cell death in an NLRP3 inflammasome-dependent manner. | Nature Communications

Figure 6: 25-HC promotes the microglial recruitment and oligodendrocyte cell death in an NLRP3 inflammasome-dependent manner.

From: 25-hydroxycholesterol contributes to cerebral inflammation of X-linked adrenoleukodystrophy through activation of the NLRP3 inflammasome

Figure 6

(a) Immunohistochemical staining of coronal brain sections with anti-Iba-1 antibody (green) and DAPI (blue) after bilateral stereotaxic injection of 25-HC (100 μM) or vehicle into the brain of Nlrp3+/+ or Nlrp3−/− mice. Scale bar, 200 μm. (b) Quantification of Iba-1+ microglia in confocal images of vehicle- or 25-HC-injected brains from Nlrp3+/+ (WT) or Nlrp3−/− (KO) mice. (n=6). (c) Quantification of IL-1β in the brain homogenates of vehicle- or 25-HC-injected hemispheres from Nlrp3+/+ (WT) or Nlrp3−/− (KO) mice. (n=7) (d) Immunohistochemical staining of brain sections with anti-Iba-1 antibody (green) after stereotaxic injection of 25-HC (100 μM) together with recombinant IL-1 receptor antagonist (IL-1Ra, 5 μg). Scale bar, 50 μm. (e) Quantification of Iba-1+ microglia in the confocal images of vehicle- or 25-HC-injected brain sections as treated in d. (n=4) (f) Immunohistochemical staining of brain sections with anti-active caspase-3 antibody (green), anti-myelin basic protein antibody (MBP, red), and DAPI (blue). Lower panel is representative magnified image of a 25-HC-injected region. Arrows indicate the active caspase-3-positive oligodendrocytes (yellow). Scale bars, 200 μm (upper); 50 μm (lower). (g) Immunohistochemical staining of brain sections of Nlrp3+/+ or Nlrp3−/− mice stained as in f after stereotaxic injection of vehicle or 25-HC. Scale bar, 100 μm. (h) Quantification of active caspase-3+ oligodendrocytes in the confocal images of vehicle- or 25-HC-injected brain sections as treated in g. (n=6) (i) Quantification of active caspase-3+ oligodendrocytes in the confocal images of vehicle- or 25-HC-injected brain sections of Abcd1+/+ (WT) or Abcd1−/− mice. (n=6) For all panels, *P<0.05, **P<0.01 and ***P<0.001.

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