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. 2021 Jul 1;12:698185. doi: 10.3389/fphys.2021.698185

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Copyright © 2021 Fan, Jiang, Gao, Bigalke, Chen, Yu, Chen and Shan.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Orexin-A treatment increases CaMKII subunits’ protein expression as well as CaMKII activation in PC12-OX1R cells. PC12-OX1R cells were incubated with 100 nM orexin-A for the indicated periods. Proteins were separated and probed with CaMKIIa, CaMKIIb, P-CamKII, and β-actin antibodies. (A) Orexin-A treatment increases CaMKII protein expression in a time-dependent manner. (A1) Representative western blots showing CaMKIIa, CaMKIIb, and β-actin in orexin-A treated and control cells. Quantification of CaMKIIa (A2) and CaMKIIb (A3) that has been normalized against β-actin and compared with control, normalized to unity (n = 6, *P < 0.05). (B) Orexin-A treatment results in a time-dependent increase in P-CaMKII level. (B1) representative western blots showing P-CaMKII and β-actin. (B2) Quantification of P-CaMKII that has been normalized against β-actin and compared with control, normalized to unity (n = 6, *P < 0.05, **P < 0.01).