Journal: Nature Communications

Article Title: Breast cancer remodels lymphatics in sentinel lymph nodes

doi: 10.1038/s41467-025-64981-z

Figure Lengend Snippet: a Expression of MGP in LECs after direct exposure to recombinant VEGF165 (5, 25, or 100 ng/mL), VEGF-C (5, 50, or 100 ng/mL), TGF-β (5, 25, or 100 ng/mL), or EGF (5, 25, or 100 ng/mL) are shown ( n = 4 HLECs). Data were depicted as Tukey box plots and analyzed using one-way ANOVA linear mixed models (Sidak correction) fitted separately for each parameter with group (recombinant vs control), dose and their interaction as fixed effects. b Gene expression changes in modified CM-exposed LECs are shown as determined by qPCR. CM was generated in the presence of antibodies against VEGFR3 (1 or 10 μg/mL, n = 4 HLECs), TGF-β (2 or 20 μg/mL, n = 8–10 HLECs), and EGF (1 or 10 μg/mL, n = 3–5 HLECs), compared to isotype control exposed samples and data were depicted as Tukey box plots showing relative gene changes and analyzed using a one-way ANOVA linear mixed models (Sidak) fitted separately for each parameter with group (antibody vs control) and dose and their interaction as fixed effects. c MGP expression determined by qPCR. CM generated with culture media devoid of VEGF supplement was used. Data were shown as Tukey box plots ( n = 9 HLECs). Data were analyzed using a one-way ANOVA linear mixed model (Sidak). The center line of the box plots represents the median, the box the 25th to 75th percentiles and the whiskers the inner fences. Statistics of group and dose effects are presented within the boxes; significant differences in comparison to the controls (defined as 1) are indicated by p values. Source data, non-significant p values and detailed experiment and n -numbers (biologically independent samples of cultured cells) are provided in the Source Data file.

Article Snippet: T47D breast cancer cells (50,000 cells) were incubated on ice for 30 min with or without 5 μg/ml recombinant MGP (His) protein (MedChemExpress P702847) and with anti-His-PE antibody (1:25; BioLegend, 362603) in RPMI + 2% FCS.

Techniques: Expressing, Recombinant, Control, Gene Expression, Modification, Generated, Comparison, Cell Culture

Journal: Nature Communications

Article Title: Breast cancer remodels lymphatics in sentinel lymph nodes

doi: 10.1038/s41467-025-64981-z

Figure Lengend Snippet: a MGP expression in LECs after CM treatment with anti-MGP antibody ( n = 5–6 HLECs), analyzed using one-way ANOVA linear mixed models (Sidak correction). b MGP expression in LECs exposed to anti-MGP antibody or recombinant MGP ( n = 6–10 HLECs), analyzed using two-way ANOVA linear mixed models (Sidak correction). c MGP expression in siRNA-silenced LECs ( n = 5 HLECs), analyzed by two-sided Mann–Whitney U -test. d Tube formation quantified by number of nodes, junctions and branches in MGP-silenced vs. control LECs, shown as geometric mean with 95% CI ( n = 17 HLECs), analyzed with a two-sided paired t -test. e Scratch assay of MGP-silenced and control LECs over 2 days (mean ± SEM; n = 13 HLECs), analyzed by repeated measures two-way ANOVA (matched full mixed model) with Sidak’s multiple comparisons. f Soluble MGP binding to cancer cells: MGP (His) was added with anti-His antibody (red), while the control (blue) contained only cancer cells with antibody. Histograms represent two independent experiments. g Ex vivo adhesion assay of T47D cells binding to lymphatic sinuses of six metastatic and five non-metastatic LNs from four and five patients, respectively (two-sided ratio paired t -test). LN sections were treated with anti-MGP or control antibody. The binding after control antibody was defined as 100% due to day-to-day variation. Example images show T47D cells binding after control vs. anti-MGP treatment to the same metastatic LN area. Adherent cells (some marked by yellow arrowheads) lie on top of tissue sections, focus adjusted to highlight cell adhesion. Scale bar: 50 μm. For box plots: center line = median; box = 25th–75th percentiles; whiskers = inner fences. Source data, non-significant p values and detailed experiment and n -numbers (biologically independent samples of cultured cells) are provided in the Source Data file.

Article Snippet: T47D breast cancer cells (50,000 cells) were incubated on ice for 30 min with or without 5 μg/ml recombinant MGP (His) protein (MedChemExpress P702847) and with anti-His-PE antibody (1:25; BioLegend, 362603) in RPMI + 2% FCS.

Techniques: Expressing, Recombinant, MANN-WHITNEY, Control, Wound Healing Assay, Binding Assay, Ex Vivo, Cell Adhesion Assay, Cell Culture

Journal: bioRxiv

Article Title: A Tough Biointerface in Human Knee Empowered by Dynamic Phase-transforming Minerals in Collagenous Matrix

doi: 10.1101/2024.08.03.606023

Figure Lengend Snippet: ( A ) Schematic illustration of the experimental procedure of proteomic analysis. ( B ) Classification of interface-enriched proteins based on functions described in Genecards Database. ( C ) Volcano plots of protein expression differences between interface and bone, and interface and root. Proteins with statistically significant differences in expression were colored in orange (significantly higher in interface) and blue (significantly lower in interface) ( p adjusted value < 0.05, fold change > 2). ( D ) Immunofluorescent staining of MGP at the S-H interface. ( E ) MGP protein expression profile across the S-H interface quantified using fluorescent signal intensity which increased in the mineralized tissue region and S-H interface (denoted by dashed line). ( F ) Functional verification of MGP’s role as a mineralization inhibitor through in vitro mineralization using nano-ACP. TEM images showed the increase in HAP crystals in the non-treated group was less than in the MGP-treated group on day 1 and day 3. Inlets revealed diffraction patterns of associated areas. ( G ) Quantification of HAP crystal density in TEM snapshots (n=5) of MGP-treated and non-treated groups. Statistical significance is illustrated as: *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001. Abbreviations: LC-MS/MS, liquid chromatography−tandem mass spectrometry; MGP, matrix Gla protein.

Article Snippet: 200-mesh golden grids with a carbon support film were added with 3 μL recombinant matrix gla protein (MGP) (100 μg/mL, CUSABIO, China) diluted with deionized water and the control group was added with 3 μL deionized water, and both were incubated for 20 minutes at room temperature.

Techniques: Expressing, Staining, Functional Assay, In Vitro, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Mass Spectrometry

Journal: bioRxiv

Article Title: A Tough Biointerface in Human Knee Empowered by Dynamic Phase-transforming Minerals in Collagenous Matrix

doi: 10.1101/2024.08.03.606023

Figure Lengend Snippet: The left panel illustrates the hierarchical structure of the meniscus root-bone interface under static condition. From macroscale to nanoscale: histological zones of the root-bone interface; transformation of mineral aggregates from ACP, immature HAP to HAP alongside mineralization of collagen fibrils; ACP, immature HAP and collagen form molecular bonds with collagen fibrils, while MGP protein and proteoglycans interact with the mineral aggregates and collagen to mediate mineralization. The right panel demonstrates the multiscale mechanical response upon mechanical loading. From macroscale to nanoscale: soft tissue responds to stress first via fiber stretching and straightening while the interface responds last, and cracks would be deflected or blunted; the ACP and immature aggregates dissipate stress (denoted by “E”) through sliding in the interfibrillar space, and crack would be arrested by stiff mineral aggregates; bonds among mineral aggregates and collagen could break for energy dissipation, and new bonds formation could occur simultaneously. Abbreviations: LR, ligamentous root; FC, fibrocartilage; MFC; mineralized fibrocartilage; ACP, amorphous calcium phosphate; HAP, hydroxyapatite; MGP, matrix gla protein.

Article Snippet: 200-mesh golden grids with a carbon support film were added with 3 μL recombinant matrix gla protein (MGP) (100 μg/mL, CUSABIO, China) diluted with deionized water and the control group was added with 3 μL deionized water, and both were incubated for 20 minutes at room temperature.

Techniques: Transformation Assay