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. 2020 Feb 4;11:85. doi: 10.3389/fimmu.2020.00085

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Copyright © 2020 Casanova, Sousa, Shakamuri, Svoboda, Buch, D'Acremont, Christophorou, Pohl, Stevens and Barlow.

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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Human rhinovirus and Poly I:C stimulation increase protein citrullination in 16HBE14°− cells. Human bronchial epithelial (16HBE14°−) cells were infected with HRV1B MOI = 5 or treated with Poly IC and total citrullination was measured with a monoclonal antibody against peptidyl citrulline (F95). (A) After 24 h, lysates were obtained and blotted with F95 antibody, α-citrullinated histone H3, and actin, which was used as loading control. (B) Confocal microscopy images were also taken at 24 or 48 h after infection, showing F95 staining (RED) and DAPI as nuclear counterstaining (BLUE). (C) Quantification of at least four different fields of view from (C) is shown in (D), with each dot representing a different cell and boxes displaying min to max and mean values of F95 staining intensity.