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. 2013 Mar 28;9(3):e1003272. doi: 10.1371/journal.ppat.1003272

Figure 4. ANDV nucleocapsid protein inhibits granzyme B-activity.

Figure 4

Analysis of possible interactions between ANDV nucleocapsid protein and granzyme B. (A) Western blot analyses showing cleaved fragments of the ANDV nucleocapsid protein (ANDV-N) after incubation with recombinant granzyme B. Full-length and cleaved ANDV-N was visualized with the mAb 7B3/F7. One representative experiment out of three is shown. (B) Granzyme B-activity was measured after pre-incubation of recombinant granzyme B with recombinant ANDV nucleocapsid protein (rANDV-N) for 30 minutes. Results were compared to granzyme B-activity for granzyme B pre-incubated with control protein (rDHFR). Activity after co-incubation with rDHFRS represents maximal granzyme B-activity. Data shown are mean ± SEM of three independent experiments carried out in duplicate. Two-tailed Student's t test was used for statistical evaluation; *** p<0.001. (C) Granzyme B-activity after pre-incubation for 30 minutes with increasing amount of recombinant ANDV nucleocapsid protein (rANDV-N). Data shown are mean ± SEM of three independent experiments carried out in duplicate. One-way ANOVA was used for statistical evaluation: except for 1 µg versus 0.75 µg (non significant), p<0.001 for all other comparisons. (D) Granzyme B-activity after pre-incubation for 0.5–27 hours of recombinant granzyme B with recombinant ANDV nucleocapsid protein (rANDV-N) compared to control protein (rDHFR). Data shown are from one experiment carried out in duplicate.