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. 2017 Jun 28;7:4340. doi: 10.1038/s41598-017-04292-6

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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P2Y₂ and K_Ca3.1 protein expression and electric response to agonists in esiRNA transfected SKOV-3 cells. (A) Analysis by immunocytochemistry after 48 h of esiRNA treatment in control (CNT) and P2Y₂-esiRNA-treated groups. Panels show the fluorescence signal for DAPI (blue) in the first column, the signal obtained with a specific antibody against P2Y₂ receptor protein (in green) in the second column, and the corresponding merged image. (B) Similar analysis was made in K_Ca3.1-esiRNA-treated cells using an antibody against K_Ca3.1 channel protein (in A and B bar = 50 µm). (C) Traces illustrate current response elicited by agonists (all 3 µM) or 1-EBIO (300 µM) in SKOV-3 cells 24 h after esiRNA transfection. In (D and E) traces illustrate current responses in SKOV-3 cells 24 h and 72 h after esiRNA transfection, respectively. Currents were normalized against the ATP-elicited response in the CNT group (18–19 cells in each condition). Every set of bars corresponds to either CNT, P2Y₂-esiRNA, or K_Ca3.1-esiRNA as indicated in the group for ATP application (*p < 0.05, experimental condition vs. respective CNT).