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Fig. 1 | Journal of Neuroinflammation

Fig. 1

From: Microglial VPS35 deficiency regulates microglial polarization and decreases ischemic stroke-induced damage in the cortex

Fig. 1

Decreases of infarct volume and neuronal death in microglial VPS35 CKO cortex in response to ischemia/stroke. a Schematic of the time course for stamoxifen injection and ischemic stroke injury in microglia-specific VPS35 conditional knockout (CKO) mice (VPS35CX3CR1). Tamoxifen (100 mg/kg) was injected (i.p.) into VPS35CX3CR1 and control (CX3CR1-CreER) mice at P15. Photothrombotic stroke was induced at P50, and mice were sacrificed 3 days after stroke. Their brain sections were subjected to TTC (2,3,5-triphenyltetrazolium chloride) staining (c, d), Nissl staining (e, f), or co-immunostaining analyses (g, h). The homogenates of the cortex were subjected to Western blot analysis (i–k). b Illustration of the photothrombotic stroke method. The mouse was injected (i.p.) with Rose Bengal (0.1 mg/g body weight), and 10 min later, its skull at the region of somatosensory cortex was exposed to a cold light to induce ischemic infarct. The ischemic core, peri-infarct, and contralateral cortex are indicated. c Representative images of TTC staining analysis. Bar, 5 mm. d Quantitative analysis of ischemic injury volumes. e, f Representative images of Nissl staining and quantitative analyses of injured brain regions. Bar, 500 μm. g, h Representative images of co-immunostaining analysis with antibodies against cleaved caspase 3 (a cell death marker) and NeuN (a neuronal marker) (g). Bar, 50 μm. Quantitative analysis (mean ± SEM; n = 5/group; *P < 0.05) was included (h). ik The homogenates from contralateral (con) and ipsilateral (ipsi) cortical brains were subjected to Western blot analysis using indicated antibodies (i) and quantification analyses (j, k). Note that the membrane was blotted with the anti-Caspase 3 first, and then reblotted with anti-β-actin. l Mouse behavior tests to access the brain injury–associated deficits. The modified neurological severity score (mNSS) test, adhesive removal, and foot fault assays were described in the “Methods” section. Data presented are mean ± SEM; n = 10/group; *P < 0.05

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