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. 2017 Apr 21;8:692. doi: 10.3389/fmicb.2017.00692

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Copyright © 2017 Cheng, Yang, Wang, Lin and Chen.

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) or licensor 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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Measurement of PB1-F2 half-life. Plasmids expressing FLAG-PB1-F2 (HK156) and FLAG-PB1-F2 (PR8) were transfected into HEK293T cells for 16 h before cycloheximide was added. Cells were harvested at 0, 0.5, 1, 2, 4, and 8 h post-cycloheximide treatment for Western analysis using anti-FLAG antibody. Additional three sets of experiments were performed without 0.5 h time point. (A) Representative blots for PB1-F2 (HK156) (Top) and PB1-F2 (PR8) (Bottom) were shown. V, vector control. (B) Chemiluminescence intensity for bands specific to PB1-F2 at each time point were captured and measured by a biomolecular imager and plotted after normalized with β-actin. Band intensity at time zero was set as 1. Regression formulation calculated from three sets of experiment is y = –0.103x+0.7205 for PB1-F2 (HK156) and y = –0.0799x+1.0031 for PB1-F2 (PR8). Protein half-life (t1/2) for each PB1-F2 was as indicated.