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. 2019 Jun 11;10:1323. doi: 10.3389/fimmu.2019.01323

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Copyright © 2019 Kimura, Mutoh, Hisamoto, Saito, Takahashi, Asakura, Ishii, Nakamura, Iida, Hase and Iwanaga.

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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RANKL induced GP2⁺Tnfaip2⁺ cells in the tracheal epithelial culture. (A) Harvested tracheal epithelial cells were cultured with 5 μg/mL GST or GST–RANKL for 6 days under ALI conditions. Immunofluorescence images of GP2 (magenta) and Tnfaip2 (green) are shown. Nuclei were stained with DAPI (blue). Bars: 100 μm. (B) Quantification of the area of Tnfaip2-positive cells. Data were obtained from 33 area in each group from 3 independent experiments and expressed as mean ± standard deviation. ***P < 0.005 calculated with Student's t-test; (C) Orthogonal images of Tnfaip2 (green), GP2 (magenta), F-actin (gray), and nuclei (blue) from confocal laser scanning microscopy. Bar: 20 μm. (D) Immunofluorescence images of Spi-B (magenta), Tnfaip2 (green), and GP2 (cyan). Nuclei were stained with DAPI (blue). Bars: 20 μm. Images are representative of at least three independent experiments.