Journal: Advanced Science

Article Title: Extracellular Vesicles Derived from Ligament Tissue Transport Interleukin‐17A to Mediate Ligament‐To‐Bone Crosstalk in Ankylosing Spondylitis

doi: 10.1002/advs.202406876

Figure Lengend Snippet: Pathological ossification and inflammatory activation in non‐ossified ligaments from patients with AS. A). µCT images show the 3D reconstruction and spinal ligament tissue from patients with AS. The red pentagram indicates uncalcified ligament. B). SOFG and HE staining of the AS patient ligament sections showing osteogenic differentiation and disorganized arrangement of collagen fibers. C,D). Representative immunohistochemistry and immunofluorescence staining images showing over‐expression of RUNX2, CD68, and PDGFRα in AS spinal ligament tissue. E). Heatmap of differentially upregulated genes, regulation of osteoblast differentiation (GO:0045667), and cytokine‐mediated signaling pathway (GO:0019221). F). Quantitative analysis of the immunofluorescence area occupied by stained regions in the different groups in (D). G). Concentration of CXCL12 in AS ligament tissue measured using ELISA. H). Pearson's correlation analysis of PDGFRα expression and the expression of markers of AS‐related osteoblast differentiation (RUNX2 and CXCL12) and inflammation (CD68) based on the ELISA and immunofluorescence staining results. I). Left: experimental scheme of AS or HD ligament tissue co‐culture with AS‐MSCs. Middle: ARS staining of AS‐MSCs after 21 days of osteogenic induction. Right: quantitative analysis of the ARS staining (absorption, 562 nm). n = 5 per group. J). Experimental scheme of AS or HD ligament tissue co‐culture with AS‐BMMCs. Concentration of inflammation factors after co‐culturing AS or HD ligament tissue with AS‐BMMCs measured using ELISA. IL, interspinous ligament; SL, supraspinous ligament; SP, spinous process; HE, hematoxylin‐eosin staining; SOFG, Safranin O–Fast Green staining.

Article Snippet: The following ELISA kits were used according to the manufacturer's protocol: CXCL12 (Solarbio, China), TNF‐α (Solarbio, China), IL‐1β (Solarbio, China), IL‐6 (Solarbio, China), and MMP14 (Thermo Scientific, USA).

Techniques: Activation Assay, Staining, Immunohistochemistry, Immunofluorescence, Over Expression, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing, Co-Culture Assay

Journal: Advanced Science

Article Title: Extracellular Vesicles Derived from Ligament Tissue Transport Interleukin‐17A to Mediate Ligament‐To‐Bone Crosstalk in Ankylosing Spondylitis

doi: 10.1002/advs.202406876

Figure Lengend Snippet: AS‐LTEVs facilitate inflammation activation by delivering IL‐17A in vivo. A). Diagrammatic representation of experimental strategies for macrophage polarization. B). Representative flow cytometry results of M1 macrophages after treatment with PBS, HD‐LTEVs, AS‐LTEVs, AS‐LTEVs with secukinumab, AS‐LTEVs with secukinumab, and Dynasore. Percentages indicate the proportion of M1 macrophages. n = 3 per group. C). Representative flow cytometry results of DCFH‐DA after treatment with PBS, HD‐LTEVs, AS‐LTEVs, AS‐LTEVs with secukinumab, AS‐LTEVs with secukinumab, and Dynasore. The right panel shows the quantitative analysis of the positive cell rate of ROS in the different groups. n = 4 per group. D–F). ELISA‐based measurement of TNF‐α, IL‐10, and IL‐1β levels in supernatants prepared from the culture medium after each of the different treatments in (B). n = 5 per group. G). Schematic diagram of the experimental schedule for the CAIA mouse model. H). µCT scans showing pathological new bone formation in hind paws in different CAIA model mice groups. I). HE staining of CAIA model mice paws and ankle joints sections showing articular destruction and inflammatory cell infiltration. J). Quantitative analysis of structural parameters of new bone by µCT analysis. n = 5 per group. K–M). Concentration of IL‐17A, TNF‐α, and IL‐6 in the serum of CAIA model mice. N. Therapeutic efficacy of the various substances assessed in terms of changes in the main symptoms of CAIA: arthritis clinical score, paw thickness, paw temperature, and pain‐associated behavior analysis using the von Frey filament test. n = 5 per group. DCFH‐DA, dichlorodihydrofluorescein diacetate; ROS, reactive oxygen species.

Article Snippet: The following ELISA kits were used according to the manufacturer's protocol: CXCL12 (Solarbio, China), TNF‐α (Solarbio, China), IL‐1β (Solarbio, China), IL‐6 (Solarbio, China), and MMP14 (Thermo Scientific, USA).

Techniques: Activation Assay, In Vivo, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Concentration Assay, Drug discovery

Journal: The EMBO Journal

Article Title: TRPML1 suppresses pulmonary fibrosis by limiting collagen and elastin deposition

doi: 10.1038/s44318-026-00712-4

Figure Lengend Snippet: ( A ) UMAP plot showing annotated cell clusters. Major cell types are labeled and color-coded, including epithelial, immune, endothelial, and stromal populations. MMP expression was determined in 32 different cell types. ( B ) UMAP plots showing the expression of selected MMP genes (MMP10, 11, 15, 16, 17, 20, 21, 23, 24, 25, 27, 28) in single cells from mouse lung tissue (filtered air group, n = 9). Expression is color-coded by log-normalized expression levels, with darker shades indicating higher expression. ( C – E ) qRT-PCR data showing mRNA expression levels of Mmp1a, Mmp1b, Mmp2, Mmp3, Mmp9, Mmp12, Mmp13, Mmp14, and Mmp19 in pmLF, IMΦ or AMΦ (WT and Trpml1 −/− ). ( F ) qRT-PCR data showing mRNA expression levels of Timp1, Timp2, Timp3, Timp4 in pmLF and IMΦ (WT and Trpml1 −/− ). In all figures, each single dot corresponds to one biologically independent sample. Data were mean ± SEM. Statistical analysis for qRT-PCR data were performed with multiple t -test, corrected for multiple comparisons using the Holm–Šídák method. ( G – J ) Western Blot analysis of different MMPs in pmLF isolated from WT and Trpml1 −/− mice. Graphs show quantification of each MMP band normalized to ß-actin. Each single dot corresponds to cells isolated from one mouse. Data were mean ± SEM. Student’s t -test, unpaired, two-tailed. .

Article Snippet: MMP14 ELISA kit , Novus biologicals , NBP3-06941.

Techniques: Labeling, Expressing, Quantitative RT-PCR, Western Blot, Isolation, Two Tailed Test

Journal: The EMBO Journal

Article Title: TRPML1 suppresses pulmonary fibrosis by limiting collagen and elastin deposition

doi: 10.1038/s44318-026-00712-4

Figure Lengend Snippet: ( A ) UMAP plot showing annotated cell clusters. Major cell types are labeled and color-coded, including epithelial, immune, endothelial, and stromal populations. MMP expression was determined in 32 different cell types. ( B ) UMAP plots showing the expression of selected MMP genes (MMP10, 11, 15, 16, 17, 20, 21, 23, 24, 25, 27, 28) in single cells from mouse lung tissue (filtered air group, n = 9). Expression is color-coded by log-normalized expression levels, with darker shades indicating higher expression. ( C – E ) qRT-PCR data showing mRNA expression levels of Mmp1a, Mmp1b, Mmp2, Mmp3, Mmp9, Mmp12, Mmp13, Mmp14, and Mmp19 in pmLF, IMΦ or AMΦ (WT and Trpml1 −/− ). ( F ) qRT-PCR data showing mRNA expression levels of Timp1, Timp2, Timp3, Timp4 in pmLF and IMΦ (WT and Trpml1 −/− ). In all figures, each single dot corresponds to one biologically independent sample. Data were mean ± SEM. Statistical analysis for qRT-PCR data were performed with multiple t -test, corrected for multiple comparisons using the Holm–Šídák method. ( G – J ) Western Blot analysis of different MMPs in pmLF isolated from WT and Trpml1 −/− mice. Graphs show quantification of each MMP band normalized to ß-actin. Each single dot corresponds to cells isolated from one mouse. Data were mean ± SEM. Student’s t -test, unpaired, two-tailed. .

Article Snippet: The following ELISAs were used for the experiments: SP-A ELISA (NBP2-76693, Novus biologicals), Cathepsin K ELISA (NBP3-00426, Novus biologicals), TIMP1 ELISA (ab196265, Abcam), TIMP2 ELISA (ab227893, Abcam), MMP1 ELISA (NBP3-06885, Novus biologicals and ABIN6963621, Antibodies online), MMP2 ELISA (ab254516, Abcam), MMP3 ELISA (ab100731), MMP8 ELISA (ab206982, Abcam), MMP9 ELISA (MMPT90, R&D systems), MMP12 ELISA (ab213878, Abcam), MMP13 ELISA (NBP3-06930, Novus biologicals), MMP14 ELISA (NBP3-06941, Novus biologicals and ABIN6957687, Antibodies online), MMP19 ELISA (NBP3-06941, Novus biologicals), IL-17A ELISA (ab199081, Abcam), DESMOSINE ELISA (CSB-E14196m, Cusabio).

Techniques: Labeling, Expressing, Quantitative RT-PCR, Western Blot, Isolation, Two Tailed Test