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. 2021 Aug 26;22(17):9227. doi: 10.3390/ijms22179227

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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K8/K18 are potential substrates of Akt. (A) A K8 or K18 phosphorylation-deficient mutant was transfected into BHK-21 cells, and the cells were treated with OA for 2 h. The transfected cells were immunoprecipitated against phosphorylated Akt substrate (PAS) antibodies followed by immunoblotting with K8/K18. (B) The Akt substrate motif was analyzed in K8 protein sequences by Scansite (scansite4.mit.edu). K8 T26 was shown as a potential Akt substrate. (C,D) BHK-21 cells were transfected with K8 WT, T26R, or T26A together with K18 WT and Akt WT and then cultured under basal (C) or OA treatment (D) conditions. The cells were immunoprecipitated against K8/K18, and then the immunoprecipitates were immunoblotted with PAS antibodies. Each PAS antibody detects the following consensus sequence: PAS #9614, RXXpS/pT; PAS #10001, RXRXXpS/pT.