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. 2017 Dec 15;7:17651. doi: 10.1038/s41598-017-17744-w

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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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Knockout of AMPKα1 and α2 expression in cultured ARPE-19 cells did not affect the inhibitory effect of AICAR on TNF-α-induced CFB expression. ARPE-19 cells were transfected with guide RNA-Cas9 leading to specific knockout of the AMPKα1 and α2 (gAMPKα), or transfected with a CRISPR/Cas9 vector guided by scramble RNA (NC; negative control). Both groups were incubated with 2.0 mM of AICAR starting 1 hour prior to stimulation with TNF-α (10 ng/mL) for 24 hours. The expression level of CFB and AMPK α1 and α2 were then examined by Western blot. Representative blots are shown.