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. 2020 Oct 15;11:538355. doi: 10.3389/fmicb.2020.538355

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Copyright © 2020 Zhang, Lin, Qin, Huang, Sun, Zhao and Jin.

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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Colocalization of nucleoprotein (NP), polymerase basic protein 2 (PB2), and polymerase basic protein 1 (PB1) with CCT5 protein. (A) CCT5-Flag and HA-NP, HA-PB2, HA-PB1, or HA-PA were cotransfected into DF-1 cells for 24 h. CCT5-Flag and HA-NP, HA-PB2, HA-PB1, or HA-PA were labeled with anti-Flag mouse and anti-HA rabbit antibodies and stained with antimouse 594 red fluorescent secondary antibody and 488 green fluorescent antirabbit antibodies, respectively. The nucleus was stained with 4’,6-diamidino-2-phenylindole (DAPI). The red, green, blue, and yellow fluorescence represents the CCT5-Flag, the HA-NP, HA-PB2, HA-PB1, or HA-PA, the localization of the nucleus, and the colocalization region, respectively. (B) DF-1 cells were infected with or without the H5N6-WT virus, and then, anti-CCT5 rabbit red fluorescent antibody and anti-NP/PB2 mouse green fluorescent antibody were used for immunostaining. The white boxed region was enlarged, as shown in the right. These experiments were examined with a confocal microscope (LSM 880; Zeiss) from three independent experiments.