Fig 1: Various membrane-coated NTUs regulate intracellular metabolic homeostasis.(a) Immunofluorescence staining of Col II, aggrecan, MMP13, and ADAMTS-5 in human chondrocytes (scale bar, 10 μm). Human chondrocytes were stimulated with IL-1β for 24 h followed by CM-NTU treatment for 6 h with red light irradiation (80 µmol photons m−2 s−1, 30 min). (b, c) ATP (b) and NADPH (c) levels in human chondrocytes incubated with IL-1β or IL-1β + CM-NTUs with light (n = 5, mean ± SD). CM-NTUs with light can increase intracellular ATP and NADPH levels close to control levels. (d–i) ATP (d, f, h) and NADPH (e, g, i) levels in muscle satellite cells (SCs), nucleus pulposus cells (NPCs), and HUVECs. SCs, NPCs, and HUVECs were incubated with the corresponding membrane-coated NTUs with or without light irradiation (n = 5, mean ± SD). ATP and NADPH concentrations after light exposure were increased 3.17–3.78 and 1.37–1.40 fold those of unirradiated ATP and NADPH, respectively. (j, k) Immunofluorescence staining of myogenic markers (MyoD and MyoG) in SCs (j) and ECM markers (Col II and MMP13) in NPCs (k) (scale bar, 10 μm). SCs or NPCs were stimulated with IL-1β for 24 h followed by corresponding membrane-coated NTU treatment for 6 h with red light irradiation. (l) Immunofluorescence staining of an antioxidant marker (Nrf2) in HUVECs (scale bar, 10 μm). HUVECs were stimulated with H2O2 for 24 h followed by HUVEC membrane-coated NTU treatment for 6 h with red light irradiation. n represents the number of biologically independent samples. P values are indicated in graphs and were determined using one-way ANOVA (b–i). Source data
Fig 2: CM-NTUs improve cell anabolism.a, ATP levels of chondrocytes treated with CM-NTUs and red light irradiation (80 µmol photons m-2 s-1) for different time intervals (n = 5, mean ± s.d.). b, ATP levels of chondrocytes treated with CM-NTUs and red light irradiation for 30 min under different light intensities (n = 5, mean ± s.d.). c, NADPH levels of chondrocytes treated with CM-NTUs with different encapsulated ferredoxin (FDX) concentrations (n = 5, mean ± s.d.). d, Immunofluorescence staining (top) and quantification (bottom) of Col II, aggrecan, MMP13 and ADAMTS-5 levels in chondrocytes (n = 5, mean ± s.d.). Mouse chondrocytes were stimulated with IL-1ß for 24 h followed by CM or CM-NTU treatment for 6 h with or without red light irradiation (80 µmol photons m-2 s-1, 30 min). e, PCR with reverse transcription detection of Col2a1, Acan, Sox9, Mmp3, Mmp13 and Adamts5 expression in chondrocytes incubated with IL-1ß, IL-1ß and CM, or IL-1ß and CM-NTUs in the dark or with IL-1ß and CM-NTUs in the light (n = 5, mean ± s.d.). f, MitoSOX-Red and JC-1 staining (left) and quantification (right) of chondrocytes incubated with IL-1ß, IL-1ß and CM, or IL-1ß and CM-NTUs in the dark or with IL-1ß and CM-NTUs in the light (MitoSOX-Red, red; JC-1, red and green; n = 5, mean ± s.d.). g, Western blots of the mitochondrial biogenesis markers SIRT1, PGC1a, TFAM, NRF1 and NRF2. Chondrocytes were incubated with IL-1ß or with IL-1ß and CM-NTUs in the light. Uncropped gel is in Supplementary Fig. 1d. n represents the number of biologically independent samples. P values are indicated in graphs and were determined using one-way ANOVA (a–f). Scale bars, 10 µm (d,f).Source data
Fig 3: IGF1+ stromal cells initiate endometrial decidualization. A The cell cluster of stromal cells (22480 cells) was re-clustered into seven sub-clusters visualized by UMAP. B Violin plots of representative markers for seven main sub-clusters of stromal cells. C Immunofluorescence staining of MMP11, ADAMTS5, FABP5, PLA2G2A, PRL, and IGFBP1 in the endometrium (Endometrium, n = 3) and decidua (Decidua, n = 3). Scale bar, 20 μm. D Bubble diagram showing the average expression of selected genes for Rem_SC and dRem_SC. E Primary decidual stromal cells (DSCs) were treated with the rh-IGF1 (2 ng/mL), rh-PRL (0.1 ng/mL), or vehicle for 48 h. And then the mRNA expression levels of these genes in DSCs were measured by qRT-PCR (n = 6). Data were presented as mean ± SEM and analyzed by t test. (ns, no significance; *, p < 0.05; **, p < 0.01; ***, p < 0.001). F Diagram showed that IGF1+ stromal cells initiate endometrial decidualization under regulation of progesterone and estrogen
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