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. 2016 Jan 1;37(8):2780–2794. doi: 10.1177/0271678X16675368

Figure 4.

Figure 4.

Ischemic stroke selectively triggers inhibition of SPAK/OSR1 phosphorylation and degradation of SPAK in SHR brains. (a) Plasma membrane fraction of the IL cerebral cortex of SHR brains exhibits selective reduction of pSPAK (68 kDa) expression at 6-h reperfusion post-tMCAO. A protein band of ∼45 kDa was detected in the membrane fraction of SHR brains by anti-tSPAK antibody but not by the anti-pSPAK/pOSR1 antibody (arrowhead). The band was interpreted as a putative, proteolytic cleavage product of pSPAK. Data are mean ± SD, n = 4, *p < 0.05. (b) Cytosol fractions from IL cerebral cortex of SHR brains show reduced expression of pSPAK, tSPAK, tOSR1, and increased accumulation of the putative cleaved SPAK band (∼45 kDa) undetected by the anti-pSPAK/pOSR1 antibody (arrowhead). Data are mean ± SD, n = 4, *p < 0.05. (c) Upregulation of Dnpep protease (aspartyl aminopeptidase, ∼50 kDa) was detected only in IL cortices of SHR but not in WKY rats at 6-h reperfusion post-tMCAO. Cytosol and crude plasma membrane protein fractions were prepared from the contralateral (CL) and ipsilateral (IL) cortices of WKY and SHR. Data are mean ± SD, n = 4, *p < 0.05.