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. 2021 Nov 29;11:23031. doi: 10.1038/s41598-021-02456-z

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© The Author(s) 2021

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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Disruption of LCMT1 in HEK293T cells. HEK293T cells were infected with lentiviruses encoding a gRNA targeting a region within LCMT1 exons 1 or 2. Cells were selected, and single colony screened for LCMT1 and methylated PP2A. (A) Wild-type (WT) and hypomorph (HM) lysates probed for LCMT1, methylated PP2Acα/β, total PP2Acα/β, and B55α. (B) Western blots were quantified, and individual data points are shown for LCMT1, methylated PP2Acα/β, total PP2Acα/β, and B55α intensities normalized to total protein loading. Error bars represent the standard deviation of four replicates. Mann–Whitney test was performed *p < 0.05, ns – not significant. (C) Workflow for proteomic analysis of HEK293T LCMT1 HM cells compared to WTcells. (D) Volcano graph of known PPP interactors. The log₂ fold change ratio HM/WT is plotted against the negative log₁₀ of the p-value of the ratio HM/WT. A p-value cut-off of 0.05 (-log₁₀ of 1.3) is indicated. PP2A subunits are labeled in purple, PP4 subunits are in blue, PP6 subunits are in green, and other PPP interactors are depicted as black dots.