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. 2020 May 4;16(5):e1008188. doi: 10.1371/journal.ppat.1008188

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© 2020 Li et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 1 — (A) Chicken DF1 cells were transfected with indicated the plasmids. Immunoprecipitation was carried out to detect interaction between chMyD88 and chSPOP using the anti-FLAG or anti-Myc antibody, followed by immunoblot analysis with indicated antibodies. (B) Co-immunoprecipitation of endogenous chSPOP and endogenous chMyD88. Cell lysates were immunoprecipitated by anti-SPOP or control IgG antibody, followed by immunoblot with indicated antibodies. (C) Immunofluorescence analysis of chMyD88 and chSPOP. Chicken DF1 cells transfected with FLAG-tagged chMyD88 and Myc-tagged chSPOP. The cells were fixed and incubated with anti-FLAG and anti-Myc antibodies, followed by incubation with secondary antibody. Nuclei were stained with DAPI. The colocalization of chMyD88 and chSPOP was detected by a confocal microscopy. (D) Schematic presentation of chMyD88 and its truncation mutants. FL, full-length; DD, death domain; INT, intermediate domain. TIR, Toll Toll/interleukin-1 receptor homology domain. (E) GFP-tagged chMyD88 or its truncated mutants and Myc-tagged chSPOP were co-transfected into chicken DF1 cells. Cell lysates were immunoprecipitated with anti-GFP antibody and then immunoblotted with indicted antibodies. (F) Schematic diagram of chSPOP and its truncation mutants. MATH, meprin and TRAF homology domain; BTB, bric-a-brac, tramtrack and broad complex/POZ domain; BOX, 3-box domain together with the C-terminal nuclear localization sequence. (G) GFP-tagged chSPOP or its truncated mutants and FLAG-tagged chMyD88 were co-transfected into chicken DF1 cells. Cell lysates were immunoprecipitated with anti-FLAG antibody and then immunoblotted with indicted antibodies.