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. 2017 Oct 23;13(10):e1006691. doi: 10.1371/journal.ppat.1006691

Fig 10. Codon-modified GE-rich domain of aLANA increases OVA-peptide presentation.

Fig 10

(A) Schematic description of the aLANA expression constructs expressing either the native GE (aLANA-SIIN) or a codon-modified GE (aLANA-GEm-SIIN). (B) Representative results from flow cytometry analysis of SIINFEKL presentation on 293Kb cells. Cells were stained with APC mouse anti-H2-Kb-SIINFEKL complex (25-D1.16) 48h after transfection with the indicated constructs. (C) Percent of eGFP+ cells expressing H2-Kb-SIINFEKL complexes at the cell surface based on the analysis in B. Bars show mean ± S.D. (n = 3). (D) DNA immunization experimental layout. Mice received 20 μg of aLANA-SIIN or aLANA-GEm-SIIN plasmids in each tibial cranial muscle for electroporation. (E) Representative results from flow cytometry analysis of SIINFEKL-specific CD8+ T cells in peripheral blood at day 14 and in spleen at day 30 after the 2nd immunization. Gated CD8+ T cells are shown. (F) Kinetics of SIINFEKL-specific CD8+ T cells in peripheral blood of immunized mice. (G) Percent of SIINFEKL-specific CD8+ T cells in spleen at day 30 after the 2nd immunization. Statistical analyses by unpaired Student t-test (C and G, *p≤0.05) or two-way ANOVA and Sidak’s post-test (F, n = 4 to 6, *p≤0.05, **p≤0.01, ***p≤0.001).