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. 2023 Dec 20;625(7993):195–203. doi: 10.1038/s41586-023-06870-3

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 1 — a. Construct schematic. ARPP19 constructs phosphorylated as indicated. Most highly conserved FAM122A residues are shown. b. Schematic describing the production of PP2A:B55 and PP2A:B55-inhibitors for structural and biophysical studies. c. Immunoblots of purified PP2A:B55 (above, SDS-PAGE), with and without NaOH treatment, using antibodies detecting methylated PP2Ac (BioLegend, Cat# 828801), PP2Ac (Millipore, Cat# MABE1783) and B55 (Cell Signaling Technologies, Cat# 2290 S). d. Size exclusion chromatography (SEC) chromatogram of PP2A:B55; peak 1 corresponds to PP2A:B55; peak 2 is excess free B55. e. PP2A:B55 activity assay using DiFMUP as a substrate; PP2A:B55 concentrations 0.3125-20 nM; 2.5 nM concentration highlighted in red. f. Same as e (2.5 nM concentration) with and without the PPP inhibitor microcystin-LR (3.75 nM concentration). g. SDS-PAGE of the PP2A:B55-tpARPP19 used for Cryo-EM grid preparation and data collection. h. SDS-PAGE of the PP2A:B55-FAM122A used for Cryo-EM grid preparation and data collection. Results (c-d and g-h) representative of 3 independent experiments.