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. 2019 Apr 23;9:6394. doi: 10.1038/s41598-019-42767-w

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Phosphorylation independent interaction of Pin1 with TAZ in vitro and in vivo. (A) Pin1 interacts with S/T-P mutant of TAZ in vitro, all (six) S or T sites followed by Proline of TAZ (S/T-P) were mutated to alanines (TAZ-6A). 200 µg of cell lysates from FLAG-tagged-TAZ-WT/TAZ-6A plasmids transfected HEK293 cells were precleared with 50% GSB beads overnight at 4 °C and GST pull down assay was carried out as described using GST or Pin1-GST. (B) Pin1 interacts with S/T-P mutant of TAZ in vivo. Co-IP was carried out as described above. In brief, FLAG-tagged-TAZ-WT/TAZ-6A were transfected alone or together with HA-tagged Pin1-WT into HEK293 cells and cells were harvested in 1% NP 40 lysis buffer after 48 hrs of transfection. After checking the expression, equal amount of lysates were subjected to co-immunoprecipitation. (C) Inhibition of phosphorylation of TAZ did not abolish the interaction of Pin1 with TAZ, GST-pull down was carried out using the FLAG-tagged-TAZ transfected HEK293 cell lysates treated or not treated with calf intestinal phosphatase (CIP) and Pin1-GST.