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. 2020 Jan 22;8:3. doi: 10.1038/s41413-019-0079-2

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© The Author(s) 2020

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. 4 — Analysis of the effect of transduction of indicated TAT-fused peptides on the formation of sealing rings in osteoclasts from wildtype (WT) and L-plastin knockout (LPL−/−) mice. a Immunoblotting analysis with an antibodyto LPL. WT and LPL^−/− osteoclasts treated with bone particles and TNF-α were transduced with indicated TAT-fused FL-LPL (lane 2) or TAT-HA (lanes 1 and 3) peptide. Immunoblotting analysis with an antibody to LPL demonstrated endogenous LPL protein in WT osteoclasts (~68–70 kDa; indicated by asterisks) and transduced FL-LPL peptide in LPL^−/− osteoclasts (~75–78 kDa; lane 2). Immunoblotting with a GAPDH antibody was used as loading control. b–i Osteoclasts transduced with indicated TAT-fused peptides were plated on dentine slices for 14 h–16 h in the presence of TNF-α to determine the formation of sealing rings (b–d). Confocal images of osteoclasts are shown. Merged (red and green) images are shown in the top panels. Dentine is shown in green color (pseudocolor; middle panels). An asterisk in b and d points to resorbed area underneath the osteoclast. Actin-stained cells are shown in the bottom panels. Arrows point to sealing rings and arrowheads point to NSZs. Wavy arrows point to podosomes. Scale bar—50 μm. Experiments were repeated three times with three different osteoclast preparations.