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. 2018 Jul 6;6:20. doi: 10.1038/s41413-018-0020-0

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© The Author(s) 2018

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. 3 — The maintenance of accumulated LD is associated with the resistance of articular chondrocytes to lipotoxicity. a Representative Nile red and BODIPY staining. Oleate at 1–1.25 mmol·L^-1 induced giant LD. The quantification of the BODIPY staining showed that 1 and 1.25 mmol·L^-1 oleate significantly increased the total LD volume (n = 4). **P < 0.01 vs. control according to Scheffe’s test. Compared to the cells treated with 1 and 1.25 mmol·L^-1 oleate, the LD size was much smaller in cells treated with 1.5 mmol·L^-1 oleate. The quantification of the BODIPY staining showed that 1.5 mmol·L^-1 oleate significantly reduced the total LD volume (n = 4). **P < 0.01 vs. 1 and 1.25 mmol·L^-1 oleate treatment according to Scheffe’s test. Scale bars, 20 μm. b Representative TUNEL, Hoechst and BODIPY triple-labelling showing that more TUNEL-positive cells are observed in cells treated with toxic concentrations of oleate (1.5 mmol·L^-1) compared to control cells or cells treated with nontoxic concentrations of oleate (1 and 1.25 mmol·L^-1). The quantification of TUNEL staining showed that 1.5 mmol·L^-1 oleate significantly increased the population of TUNEL-positive cells. **P < 0.01 vs. control and 1 and 1.25 mmol·L^-1 oleate treatment according to Scheffe’s test. The size of the LDs decreased in TUNEL-positive cells compared to TUNEL-negative cells. The quantification of BODIPY staining showed that the total LD volume was significantly decreased in TUNEL-positive cells. **P < 0.01 vs. TUNEL-negative cells according to Scheffe’s test. Scale bars, 20 μm. c Representative western blots showing that 1.5 mmol·L^-1 oleate decreased the expression level of FSP27 protein (n = 4). d The knockdown of FSP27 reduced the total LD volume. The quantification shows that the knockdown of FSP27 significantly decreased the total LD volume. **P < 0.01 vs. scRNA control according to Scheffe’s test. Scale bar, 10 μm. e Viability assay showing that FSP27 depletion significantly sensitised chondrocytes to lipotoxicity (n = 4). **P < 0.01 vs. scRNA according to Scheffe’s test. f Immunohistochemistry on cartilages obtained from ACLT model 6 weeks after surgery. The FSP27-positive cells were quantified (n = 4). The population of FSP27-positive articular chondrocytes was significantly smaller in mice fed an HFD than in mice fed an SD. **P < 0.01 vs. mice fed an SD according to Scheffe’s test. BODIPY and Hoechst dual labelling showing that LDs are accumulated in mice fed an HFD. Scale bar, 20 μm