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. 2019 Feb 15;10:771. doi: 10.1038/s41467-019-08686-0

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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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Fig. 8 — Function of the ASPP2 SH3 domain/PP1α C-tail interaction. a Quantitative AP-MS from HEK293T cells expressing Strep-HA tagged ASPP2 or ASPP2^KVK. Chart shows protein abundance normalised to the bait (%) relative to PP1α, PP1β, PP1γ1. Protein abundance is measured on the basis of the average intensity of the three most intense and unique peptide precursors. Error bars indicate the standard deviation. A two-way ANOVA test was done to determine if the differences among means were significantly different from each other. Three pairwise comparisons were carried out (ASPP2 PP1α vs ASPP2KVK PP1α, ASPP2 PP1β vs ASPP2^KVK PP1β and ASPP2 PP1ɣ1 vs ASPP2^KVK PP1ɣ1) and p-values were adjusted using Bonferroni correction. Significant differences are marked. *** indicates p < 0.0001. b Quantitative AP-MS from HEK 293T cells expressing Strep-SH tagged PP1α, PP1α^ΔC (PP1α^1-300), PP1β and PP1ɣ1. Protein abundance of ASPP1, ASPP2 and iASPP is measured on the basis of the three most intense and unique peptide precursors. Each intensity value is normalised to the respective bait and to PP1α. Error bars indicate standard deviation. A two-way ANOVA was done to determine if differences among means were significantly different from each other. Three pairwise comparison among each group were carried out (PP1ɑ vs PP1ɑΔC, PP1ɑ vs PP1β and PP1α vs PP1ɣ1) and p-values were adjusted using a Bonferroni correction. Significant differences are marked. * indicates p < 0.05, ** indicates p < 0.005. c Comparison of PP1α and PP1α^ΔC protein interactions. Heatmap of ASPP2-PP1 complex proteins intensity after purification with PP1α or PP1ɑ^ΔC. d Western blot of transfected HEK293T cell lysates probed with the indicated antibodies. e Quantification of the ratio between P-TAZ^S89 and total HA-TAZ protein levels normalised to control phosphorylation. Error bars represent standard deviation. A one-way ANOVA test was carried out to determine if the differences among means were significantly different from each other. Two pairwise comparisons were carried out (ASPP2^wt vs ASPP^FA and ASPP^wt vs ASPP^KVK) and p-values were adjusted using a Bonferroni correction. Significant differences are marked. * indicates p < 0.05 (n = 3 independent experiments)