Fig. 3

Activation of neuronal NF-κB in SHR rats following ischemia and reperfusion. (A-J) SD and SHR rats were subjected to sham surgery, or MCAO (60 min) followed by reperfusion (RPF) for 6 h. (A-H) Tissue extracts from the ipsilateral cortex were processed to measure mRNA levels of NFκB1 and RelA by quantitative real-time-PCR. Transcript levels were normalized against reference genes β-actin or HPRT. Bar graphs represent relative expression of NFκB1 and RelA genes (mean ± SE, n = 5–6). (I) Tissue lysates with equal amount of protein from the ipsilateral cortex were analyzed using anti-IκB antibody (upper panels). Equal protein loading was confirmed by re-probing the blots with anti-β-tubulin antibody (lower panel). Bar graps represent quantitative analysis of IκB protein levels (mean ± SD, n = 4). (A-I) Significant difference between means was assessed by student t-test and presented as *p < 0.05, **p < 0.01 and ***p < 0.001. (J) Coronal brain sections through the ipsilateral cortex of SD and SHR rats (MCAO 60’/RPF 6 h) were processed for immunohistochemical staining with anti-phospho-NFκB (pNFκB - red) antibody, anti-NeuN (green) antibody and nuclear stain DAPI (blue). Arrows in the representative photomicrographs demonstrate nuclear localization of phosphorylated NFκB in NeuN positive cells in the ipsilateral cortex