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. 2016 Mar 1;13:35. doi: 10.1186/s12985-016-0494-6

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© Noppakunmongkolchai et al. 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 2 — Inhibitions of PKC by PKCα siRNA or a chemical inhibitor (BisI) suppressed NS5 RdRp phosphorylations. a PKCα-specific siRNA depleted endogenous PKCα at 24, 48, 72 h after siRNA transfection. A pool of PKCα siRNAs (siPKCα) was transfected into the BHK-21 cells. The levels of PKCα were detected by PKCα immunoblotting. A non-targeting siRNA (siNeg) was used as a control. GAPDH immunoblots were used as a loading control. b PKCα depletion by siRNA downregulated NS5 Threonine and Serine phosphorylations. BHK-21 cells were infected with recombinant DENV expressing His-FLAG-tagged NS5. The His-FLAG-tagged NS5 was pulled down (IP) from BHK-21 cell lysate by anti-FLAG antibody, and Threonine/Serine phosphorylations (p-Thr/p-Ser) were examined by p-Thr or p-Ser-specific antibodies. c Quantifications of p-Thr/NS5 and p-Ser/NS5 from (b)