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. 2016 Nov 22;11(11):e0166763. doi: 10.1371/journal.pone.0166763

Fig 4. CaMKII γ and δ expressed in the Huh7.5.1 cells can directly phosphorylate NS5A S235.

Fig 4

(A) Expression of CaMKII isoforms in the Huh7.5.1 cells by RT-PCR analysis. Expression of CaMKII (γ and δ) mRNA was assessed with the genomic (g) and cDNA (c) libraries of the Huh7.5.1 cells using primers that span two neighboring exons (exon-exon junction primer) to avoid false positive signal from the genomic DNA. No template (n) served as negative controls. Lack of CaMKII (α and β) mRNA expression was double checked with primer pairs within the same exons (exon primer). The lanes are numbered. Lane M indicates DNA markers in base pairs (bp). (B) In vitro kinase assay. Three digits were used to denote the phosphorylation status of the three serine residues 222, 235 and 238 in the biotin-labeled synthetic NS5A peptide (NS5A-111 and NS5A-101). Numbers 1 and 0 represent phosphorylation and non-phosphorylation, respectively.