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. 2023 Apr 12;14:2057. doi: 10.1038/s41467-023-37632-4

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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 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. 5 — a Scheme for hGCase-hBS treatment and lysosome isolation from cells. H4 cells expressing lysosome-tag (TMEM192-3HA) were treated with 1 nM hGCase-hBS for 24 h, followed by cell lysis and lysosome isolation using anti-HA coated magnetic beads. Schematic created with BioRender.com. b Validation of lysosome enrichment after TMEM192-3HA-based Lyso-IP. Western blots showing enrichment of lysosomes after Lyso- IP as demonstrated by enrichment of bonafide lysosomal proteins LAMP2, Cathepsin D and GCase in eluent fraction compared to input. GCase is detectable in lysosomes 24 h after treatment of GBA−/− cells. Representative blots of 3 replicates. c Rescue of dysregulated lysosomal proteins upon hGCase-hBS treatment. Graph showing increased or decreased levels of lysosomal proteins in GBA−/− cells (blue bar) and its rescue upon hGCase-hBS treatment (yellow bar). d Rescue of dysregulated lysosomal lipids upon hGCase-hBS treatment. Graph showing increased or decreased levels of lysosomal lipid species in GBA−/− cells (blue bar) and its rescue upon hGCase-hBS treatment (yellow bar). Note: The lipid analysis performed does not report the number of carbons in the sphingoid base and the acyl chain (fatty acid chain) separately. Hence, the first number in the nomenclature used refers to the total number of carbons (sphingoid base + acyl chain). An Excel sheet consisting of each of the detected lipid species and its corresponding Swisslipids ID is provided in Supplementary Data File 3. FDR-corrected p-values are shown in (c) and (d) (**q-value <0.01; *q-value <0.05; n = 3 independent measurements), calculated for each contrast separately. Trends are also displayed (e.g. significance found in one contrast only), to highlight potential lipid/protein entities with opposite fold change in GBA−/− and rescue upon hGCase-hBS treatment. Source data are provided as a Source Data file. Schematics created with BioRender.com.