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. 2018 May 23;9:1114. doi: 10.3389/fimmu.2018.01114

Figure 1.

Figure 1

Recognition of recombinant human PCNA (hPCNA) by NKp44-derived pep8 (NKp44-pep8). (A) ELISA assay representing the binding curve of biotinylated NKp44-derived pep5, pep8, and pep10 at a peptide concentration range of 0–7.24 µM to plate bond hPCNA. (B) IFNγ secretion assay of NK92-44-1 cells co-incubate with the HLA positive cell line PANC-1 (E:T ration of 1:2) in the presence of NKp44-pep8 (10 or 20 μg/well), NKp44-derived pep5 was used as negative control. Note that when we apply to NK92-44-1 pep8 (without target cells), activation of IFNγ secretion is minimal and less/similar to the observed activation when applying the pep5 negative control. (C) ELISA assay of blocking NKp44–proliferating cell nuclear antigen (PCNA) interaction via NKp44-pep8. NKp44-derived pep5 was used as a negative control. No peptide represents the maximal binding capacity of NKp44 to hPCNA. (D) ELISA assay showing the specific recognition of NKp44-pep8 (5 µg/ml) to hPCNA relative to TL1A, APO-E3, HNF4, NKp44-Ig, hNKp46-Ig, hFc. (E) ProteOn array showing the binding of hPCNA at protein concentration range of 0–250 nM to bond NKp44-pep8. 0 nM (X base-line), 15.6 nM (yellow), 31.2 (purple), 62.5 nM (green), 125 nM (blue), and 250 nM (red).