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. 2020 Jun 28;19(15):1952–1968. doi: 10.1080/15384101.2020.1785724

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© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.

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Figure 5. — UAF1 and ID1 interact, and co-localize at the centrosome.

(a, b) HEK293 T cells were transfected with 3xFLAG-tagged UAF1 cDNA (FLAG-UAF1) and HA-tagged ID1 cDNA (HA-ID1). (a, b) After 48 h, whole cell extracts were prepared for immunoprecipitation (IP) with an anti-FLAG antibody (A) and an anti-HA antibody (b). Immunoprecipitates were eluted, resolved by SDS-PAGE, and immunoblotted using the indicated antibodies. (c) HEK293 T cells were transfected with a combination of UAF1 siRNA and HA-ID1 cDNA. After 48 h of transfection, whole cell extracts (WCE) were subjected for immunoblotting. (d-f) After 48 h of transfection, cells were fixed for immunostaining as indicated. HeLa_Centrin1-GFP cells (d) and HeLa cells (f) were transfected with HA-ID1 cDNA. (e) HeLa cells were transfected with mNeonGreen-fused UAF1 cDNA. Bar, 5 μm. (g) HeLa_Centrin1-GFP cells were transfected with UAF1 siRNA for 48 h and fixed. The percentage of cells with more than two centrosomes was displayed. Three independent experiments were performed. Values represent mean ± SEM. Statistics: paired t-test. *, p ≤ 0.05.