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Briefly, cells were lysed in RIPA buffer (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 1 m M EGTA, 1% NP-40, 1% sodium deoxycholate, 2.5 mM sodium pyrophosphate, 1 mM -glycerophosphate, 1 mM Na 3 VO 4 , and 1 g/ml leupeptin, 1 mM PMSF), and total cellular protein was extracted and quantitated

The cryo-EM structure of SARS-CoV-2 ORF3a is the first viroporin family structure determined in coronaviruses 142

Kalec et al

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These outcomes were accompanied by the restoration of neurotransmitter balance, particularly the enhancement of GABAergic inhibition and the reduction of excessive glutamatergic excitation