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. 2023 Feb 21;14:906. doi: 10.1038/s41467-023-36484-2

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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. 1 — a Experimental setup for SPION ‘pulse-chase’ and representative confocal images of mouse bone marrow monocyte (BMMs) differentiation into mature αvβ₃-integrin-positive (IntegriSense⁶⁴⁵) osteoclasts following RANKL stimulation. Bar 150 µm. b Quantitation of IntegriSense⁶⁴⁵ positive cells during RANKL stimulation (n = 3 biologically independent cells from 2 experiments). Data are presented as mean percentage (%) ± SD. c Schema of the SPION-based SL enrichment method, validation, and proteomic analysis. Created with BioRender.com. d Immunoblotting for protein markers of various subcellular compartments in whole-cell homogenates (H), post-nuclear supernatant (PNS), magnetic column flow-through (UB), and enriched SL fractions (F1–F3) (n = 3). LRO lysosome-related organelle. e Volcano plot depicting the difference between proteins in whole cell homogenates and secretory lysosomes (SLs) (n = 3, P > 0.05, ANOVA, Benjamini–Hochberg adjusted). The top 218 proteins up-regulated in the lysosomal fractions are shown in green (FC > 1.5, P < 0.05) with the top 6 membrane transporters colored in purple and Slc37a2 highlighted in red.