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. 2020 Sep 25;11:581867. doi: 10.3389/fmicb.2020.581867

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Copyright © 2020 Sharma, Batra, Stuchlik, Reed, Pohl, Chow, Sambhara and Lal.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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IAV NP interrupts FLNA-TRAF2 interaction, allowing for increased TRAF2-ASK1 interaction. HEK293 cells were transfected with either 5 μg of pcDNA3.1 empty control vector or NP(PR8) plasmid. The cells were harvested (A) 8 h post-transfection and (B) 24 h post-transfection and IP was setup using mouse anti-FLNA and mouse anti-TRAF2 antibody (n = 3) (see Supplementary Figures 1, 4). TRAF2 and ASK1 were detected in the eluate by Western blotting using rabbit anti-TRAF2 and mouse anti-ASK1 antibody. (C,D) Densitometric analysis was performed using the ImageJ software to visualize the level of FLNA-TRAF2 (FLNA used as loading control) and TRAF2-ASK1 (TRAF2 used as loading control) interaction. Results are shown as mean ± S.D. of one independent experiment (n = 3). Statistical significance for FLNA-TRAF2 and TRAF2-ASK1 level of interaction was determined using Student’s t test (^∗p < 0.05).