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Specifically, CHAC1 provides neuroprotection during damage induced by subarachnoid or intracerebral hemorrhage ( CHAC1/BOTCH , which significantly reduced neuroinflammation by blocking NOTCH1 signaling ( The regulation of CHAC1/BOTCH involves additional molecular players: during intracerebral hemorrhage, inhibition of CHAC1/BOTCH by the androgen receptor (AR) and Jumonji-domain containing protein-3 (JMJD3) led to elevated NOTCH1 levels and exacerbated neuroinflammation ( ATF4 was most prominently upregulated, though other related genes were also significantly elevated (Yasuda et al., 2014)

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(PubMed) May JM, Qu ZC, Mendiratta S

Next the cells were centrifuged and re-suspended in 1 mL RIP lysis buffer (150 mM KCl, 25 mM TrisHCl (pH 7.4), 5 mM EDTA, 0.5% NP-40, with 100 U/ml RNase inhibitor and protease inhibitor cocktail)