Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: Effect of EFEMP1 knockdown by small interfering RNA (siRNA) on chondrocyte phenotype. A The human chondrocyte cell line C20A4 was transfected with either EFEMP1 siRNA (siEFEMP1) or scrambled siRNA (siCtrl). EFEMP1 and MMP-13 protein expression was analyzed by Western blotting. B Chondrocyte-related gene expression, including SOX9, aggrecan (ACAN), MMP13, type X collagen (COL10A1), and type II collagen (COL2A1), was assessed by quantitative PCR in siEFEMP1- and siCtrl-treated C20A4 cells. C Phosphokinase protein arrays were used to profile phosphorylation changes in intracellular proteins extracted from C20A4 chondrocytes. D Spot intensities exhibiting changes greater than ± 25% between groups were quantified using ImageJ software. Red lines indicate baseline levels from the siCtrl group. E Phosphorylation of p70S6K at threonine 421/serine 424 (T421/T424) was further examined by Western blot analysis. Data are presented as mean ± standard error of the mean (SEM). Statistical significance was determined using an independent-samples t-test. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Knockdown, Small Interfering RNA, Transfection, Expressing, Western Blot, Gene Expression, Real-time Polymerase Chain Reaction, Phospho-proteomics, Software

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: Immunofluorescence analysis of cartilage markers following EFEMP1 antibody treatment in STR/ort mice. A Representative immunofluorescence (IF) staining for EFEMP1, SOX9, MMP13, aggrecan fragments, COL2A1, and COL10A1 in the medial femoral condyle (MF) and medial tibial plateau (MT). Nuclei were counterstained with DAPI (blue), and articular cartilage regions are outlined by orange dashed lines. B Quantification of fluorescence intensity in articular cartilage regions of the femoral condyle and tibial plateau, with medial and lateral compartments combined. EFEMP1 and SOX9 signals were normalized to DAPI, whereas MMP13, aggrecan fragments, COL10A1, and COL2A1 signals were quantified as absolute FITC intensity. Data are presented as mean ± standard error of the mean (SEM). Statistical analysis was performed using an independent-samples t -test. P < 0.05, P < 0.01, and ** P < 0.001

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Plastrum testudinis Ameliorates Oxidative Stress in Nucleus Pulposus Cells via Downregulating the TNF-α Signaling Pathway.

doi: 10.3390/ph16101482

Figure Lengend Snippet: Figure 4. Expression of ACAN and COL2A1 in NPCs. (A) Expression of ACAN and COL2A1 pro- teins in NPCs treated with TBHP, with or without PT. (B,C) Expression of ACAN and COL2A1 proteins relative to β-actin in NPCs treated with TBHP, with or without PT. (D) Representa- tive immunofluorescence images of ACAN in NPCs photographed by fluorescence microscopy (scale bar = 100 µm). (E) Representative immunofluorescence images of COL2A1 in NPCs pho- tographed by fluorescence microscopy (scale bar = 100 µm). Data are the mean ± standard deviation. #### p < 0.0001 vs. CTL group; ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. 100 µM TBHP group.

Article Snippet: ACAN (sc-166951) and COL2A1 (sc-52658) were from Santa Cruz (Shanghai, China), and prostaglandin G/H synthase 2 (PTGS2) (WL01750), endothelin-1 (EDN1) (WL02780), JUN (WL02863), FOS (WL03699), and IL-6 (WL02841) were from Wanleibio Co. Ltd. (Shenyang, China).

Techniques: Expressing, Microscopy, Standard Deviation

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Plastrum testudinis Ameliorates Oxidative Stress in Nucleus Pulposus Cells via Downregulating the TNF-α Signaling Pathway.

doi: 10.3390/ph16101482

Figure Lengend Snippet: Figure 7. Mechanism underlying the therapeutic effect of PT on IDD. PT could downregulate the expression of inflammation-specific targets including TNF-α, FOS, JUN, PTGS2 and EDN1 on the TNF-α signaling pathway as well as cellular senescence marker p16, inflammatory factor IL-6, and upregulate the expressions of COL2A1 and ACAN, thus leading to a suppressive effect on inflammatory response and oxidative stress in NPCs.

Article Snippet: ACAN (sc-166951) and COL2A1 (sc-52658) were from Santa Cruz (Shanghai, China), and prostaglandin G/H synthase 2 (PTGS2) (WL01750), endothelin-1 (EDN1) (WL02780), JUN (WL02863), FOS (WL03699), and IL-6 (WL02841) were from Wanleibio Co. Ltd. (Shenyang, China).

Techniques: Expressing, Marker

Journal: Journal of Tissue Engineering

Article Title: High BMPR2 expression leads to enhanced SMAD1/5/8 signalling and GDF6 responsiveness in human adipose-derived stem cells: implications for stem cell therapies for intervertebral disc degeneration

doi: 10.1177/2041731420919334

Figure Lengend Snippet: Immunohistochemical analysis of ASC collagen gel cultures after 14 days with 100 ng mL −1 rhGDF6 stimulation with and without SMAD1/5/8 (Dorso) or ERK1/2 (U0126) inhibition and no stimulation control. After 14 days culture, constructs stimulated with rhGDF6 demonstrated an increase in aggrecan and type II collagen deposition compared to rhGDF6 stimulated constructs where SMAD1/5/8 and ERK1/2 signalling had been blocked, where levels appeared similar to unstimulated controls. Scale bar, 500 µm.

Article Snippet: Sections were allowed to equilibrate to room temperature, formalin fixed and blocked prior to addition of either aggrecan (Bio-Rad; MCA1454G) or type II collagen (Proteintech 15943-1-AP) primary antibodies and incubated at 4°C overnight.

Techniques: Immunohistochemical staining, Inhibition, Control, Construct

Journal: Stem cell research & therapy

Article Title: Intra-renal delivery of mesenchymal stem cells attenuates myocardial injury after reversal of hypertension in porcine renovascular disease.

doi: 10.1186/scrt541

Figure Lengend Snippet: Figure 1 Hemodynamic effect of renal artery stenosis and characteristics of MSC. A) Renal angiography before and after PTRA, showing successful restoration of vessel patency (arrows). B) Mean arterial pressure (MAP) measured by telemetry decreased after PTRA. C) Immunostaining (left) and FACS (right) showing that adipose tissue-derived MSC expressed CD44 and CD90 surface markers. D) Representative immunostaining showing that MSC trans-differentiated into osteocytes, chondrocytes and adipocytes in vitro. E) DiI-labeled MSC (red) incorporated into tubes formed by HUVEC (grey). F) Representative fluorescence of CM-DiI-labeled MSC (red, arrows) and cytokeratin (green) in the stenotic-kidney (left, X20) and myocardium (right, X40) four weeks after administration. FACS, fluorescence activated cell sorting; HUVEC, human umbilical vein endothelial cells; MSC, mesenchymal stem cells; PTRA, percutaneous transluminal renal angioplasty.

Article Snippet: MSC characterization was confirmed by their trans-differentiation into osteocytes (mouse anti-human osteocalcin antibody and alizarin red staining), chondrocytes (goat anti-human aggrecan antibody) and adipocytes (goat anti-mouse FABP4 antibody and oil red staining) (R&D Systems, Pittsburgh, PA, USA) [17].

Techniques: Immunostaining, Derivative Assay, In Vitro, Labeling, Fluorescence, FACS

Journal: Oxidative medicine and cellular longevity

Article Title: Selenium Attenuates TBHP-Induced Apoptosis of Nucleus Pulposus Cells by Suppressing Mitochondrial Fission through Activating Nuclear Factor Erythroid 2-Related Factor 2.

doi: 10.1155/2022/7531788

Figure Lengend Snippet: Figure 2: Se ameliorated TBHP-induced apoptosis and ECM degeneration of NPCs. (a, b) Apoptosis of the NPCs detected by flow cytometry analysis. (c, d) Typical fluorescence photomicrograph and quantitation of TUNEL staining of NPCs (scale bar: 100 μm). (e, f) Representative western blot bands and quantitation of the expression of cleaved-caspase 9, cleaved-caspase 3, Bax, and Bcl-2 in the NPCs. (g, h) Representative western blot bands and quantitation of the expression of anabolic mediators (aggrecan and collagen II) and catabolic mediators (MMP3 and MMP9). The data are represented as the mean ± SD from at least 3 independent experiments. ∗P < 0:05 and ∗∗P < 0:01 vs. the TBHP group.

Article Snippet: After being blocked with 5% skimmed milk for 1 h at room temperature, the PVDF membranes were incubated overnight at 4°C with the primary antibodies against Nrf2 (1 : 1000, ProteinTech, Wuhan, China), HO-1 (1 : 1000, ProteinTech), SOD2 (1 : 2000, ProteinTech), dynamin-related protein 1 (DRP1) (1 : 1000, Affinity Biosciences, OH, USA), mitochondrial fission factor (MFF) (1 : 1000, ProteinTech), mitochondrial fission 1 (Fis1) (1 : 1000, ProteinTech), optic atrophy 1 (OPA1) (1 : 1000, ProteinTech), mitofusin 1 (Mfn1) (1 : 1000, ProteinTech), mitofusin 2 (Mfn2) (1 : 1000, ProteinTech), Aggrecan (1 : 500, NOVUS, USA), collagen II (1 : 1000, Affinity Biosciences), matrix metalloproteinase 3 (MMP3) (1 : 1000, Affinity Biosciences), MMP9 (1 : 1000, Affinity Biosciences), cleaved-caspase 3 (1 : 1000, Affinity Biosciences), caspase 9 (1 : 1000, ProteinTech), Bcl-2 (1 : 1000, Abcam, Cambridge, MA, USA), Bax (1 : 1000, ProteinTech), and β-actin (1 : 2000, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Cytometry, Quantitation Assay, TUNEL Assay, Staining, Western Blot, Expressing