Journal: Journal of translational medicine

Article Title: Circadian rhythm disruption upregulating Per1 in mandibular condylar chondrocytes mediating temporomandibular joint osteoarthritis via GSK3β/β-CATENIN pathway.

doi: 10.1186/s12967-024-05475-2

Figure Lengend Snippet: Fig. 2 Chronic circadian rhythm disruption leads to increased expression of the clock gene Per1 and matrix degrading enzymes in the mandibular condylar cartilage. A Relative mRNA expression curves of core clock genes in the mandibular condylar cartilage over a 24-h period (ZT0, ZT4, ZT8, ZT12, ZT16, ZT20 and ZT24) of two groups. B Western blot images of PER1, MMP13, ADAMTS4, and ADAMTS5 in mandibular condylar cartilage tissues in two groups. C Quantitative analysis of Western blot results. D Representative slices of immunohistochemical staining in the two groups. E Quantitative analysis of AOD in immunohistochemical staining of the two groups. n = 5, *P < 0.05. **P < 0.01

Article Snippet: Antigen retrieval was performed by incubating the sections in 1 mM EDTA antigen retrieval solution (pH = 9) in a water bath at 95 °C for 45 min. After rinsing with PBS, the sections were blocked with 5% BSA at room temperature for 1 h. The sections were then incubated with primary antibodies MMP13 (1:600, ProteinTech, China), ADAMTS4 (1:100, Affinity, USA), ADAMTS5 (1:100, Affinity, USA), and β-CATENIN (1:1000, ProteinTech, China) at 4 °C overnight.

Techniques: Disruption, Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Journal of translational medicine

Article Title: Circadian rhythm disruption upregulating Per1 in mandibular condylar chondrocytes mediating temporomandibular joint osteoarthritis via GSK3β/β-CATENIN pathway.

doi: 10.1186/s12967-024-05475-2

Figure Lengend Snippet: Fig. 6 Downregulation of Per1 in rat mandibular condylar cartilage can inhibit the activation of the GSK3β/β-CATENIN pathway and the increase in matrix-degrading enzymes caused by chronic circadian rhythm disruption. A Representative sections of immunohistochemical staining of PER1, MMP13, ADAMTS4, ADAMTS5 and β-CATENIN in mandibular condylar cartilage. B Average optical density of protein in condylar cartilage tissue. C Western blot results of PER1, MMP13, ADAMTS4, ADAMTS5, GSK3β, p-GSK3β, and β-CATENIN in the mandibular condylar cartilage of rats in four groups. D Statistical analysis of Western blot results in the condylar cartilage tissue of rats in the four groups. E Statistical analysis of p-GSK3β/GSK3β. n = 5, *P < 0.05, **P < 0.01

Article Snippet: Antigen retrieval was performed by incubating the sections in 1 mM EDTA antigen retrieval solution (pH = 9) in a water bath at 95 °C for 45 min. After rinsing with PBS, the sections were blocked with 5% BSA at room temperature for 1 h. The sections were then incubated with primary antibodies MMP13 (1:600, ProteinTech, China), ADAMTS4 (1:100, Affinity, USA), ADAMTS5 (1:100, Affinity, USA), and β-CATENIN (1:1000, ProteinTech, China) at 4 °C overnight.

Techniques: Activation Assay, Disruption, Immunohistochemical staining, Staining, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Physalin A Inhibits MAPK and NF-κB Signal Transduction Through Integrin αVβ3 and Exerts Chondroprotective Effect

doi: 10.3389/fphar.2021.761922

Figure Lengend Snippet: PA suppressed excess expression of the catabolic indicators of chondrocytes induced by IL-1β, including ADAMTS5, MMP1, MMP3, and MMP13. Mice chondrocytes were treated with 5 ng/ml of IL-1β, alone or with PA (2.5, 5, and 10 μM) for 24 h (A) Western blotting results and (B–E) quantitative analysis of ADAMTS5, MMP1, MMP3, and MMP13. (F) MMP13 expression was observed by immunofluorescence staining when chondrocytes were treated with 5 ng/ml of IL-1β, alone or with 10 μM of PA (scale bar 200 μm). (G–I) Relative mRNA levels of ADAMTS5, MMP3, and MMP13 in chondrocytes stimulated with 5 ng/ml of IL-1β, alone or with PA (2.5, 5, and 10 μM) for 24 h. GAPDH was used as an internal reference. Data are presented as means ± SD ( n = 3). The exact p value was marked in the corresponding figure and p < 0.05 was considered statistically significant.

Article Snippet: Primary antibody against ADAMTS5 (A02802-1) was got from Boster (Wuhan, Hubei, China) and was applied at a 1:500 dilution.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: Physalin A Inhibits MAPK and NF-κB Signal Transduction Through Integrin αVβ3 and Exerts Chondroprotective Effect

doi: 10.3389/fphar.2021.761922

Figure Lengend Snippet: Knockdown of integrin αVβ3 weakened the anti-inflammatory, anabolism enhancing, and catabolism inhibiting effect of PA on IL-1β-induced chondrocytes. (A,B) Relative mRNA levels of integrin αV (Itg αV) and integrin β3 (Itg β3) in chondrocytes stimulated with 5 ng/ml of IL-1β, alone or with PA (2.5, 5, and 10 μM) for 24 h (C,D) Itg αV and Itg β3 were knocked down by siRNA transfection, and the knockdown efficiency was verified by RT-PCR. (E,F) Inflammatory markers (COX2, iNOS) were detected by western blotting and the band density of protein levels were quantified after mice chondrocytes were added with or without 5 ng/ml of IL-1β, 10 μM of PA, and Itg αVβ3 siRNA. (G–I) Western blotting was applied to measure the anabolic (aggrecan, collagen II) and catabolic markers (MMP1, MMP3, MMP13, and ADAMTS5) in the Itg αVβ3-deficiency mice chondrocytes along with or without the administration of 5 ng/ml of IL-1β and 10 μM of PA, and the band density of these protein levels were quantified in the histogram. GAPDH was used as an internal reference. Data are presented as means ± SD ( n = 3). The exact p value was marked in the corresponding figure and p < 0.05 was considered statistically significant.

Article Snippet: Primary antibody against ADAMTS5 (A02802-1) was got from Boster (Wuhan, Hubei, China) and was applied at a 1:500 dilution.

Techniques: Knockdown, Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Physalin A Inhibits MAPK and NF-κB Signal Transduction Through Integrin αVβ3 and Exerts Chondroprotective Effect

doi: 10.3389/fphar.2021.761922

Figure Lengend Snippet: Primer sequence used in the RT-qPCR experiment.

Article Snippet: Primary antibody against ADAMTS5 (A02802-1) was got from Boster (Wuhan, Hubei, China) and was applied at a 1:500 dilution.

Techniques: Sequencing

Journal: Cancer Biology & Therapy

Article Title: Reprogramming of VEGF-mediated extracellular matrix changes through autocrine signaling

doi: 10.1080/15384047.2023.2184145

Figure Lengend Snippet: Immunoblot analysis of enzymes and receptors in cancer cells. (a) Changes in ADAMTS1, uPAR, LOX, MMP1 and MMP2 in 231_VEGF cells compared to 231_WT cells. (b) Changes in ADAMTS1, LOX, uPAR and MMP1 in wild type and VEGF overexpressing PC-3 and MCF-7 cells. (c) Changes in NRP-1 in MDA-MB-231, PC-3 and MCF-7 wild type and VEGF overexpressing cells.

Article Snippet: Antibodies cross-reactive with mouse/human ECM proteins and specific for human enzymes of interest included rabbit-polyclonal anti-Col1A1 antibody (1:1000; OriGene, Rockville, MD, USA), mouse monoclonal anti-FN1 antibody (1:2000 dilution; Proteintech, Rosemont, IL, USA), rabbit polyclonal anti-MMP-1 antibody (1:1000 dilution; Neo BioLab, Woburn, MA, USA), rabbit polyclonal anti-MMP2 (1:1000 dilution; GeneTex, Inc., Irvine, CA, USA), rabbit polyclonal anti-MMP-14 antibody (1:1000 dilution; Neo BioLab, Woburn, MA, USA), mouse monoclonal anti-lysyl oxidase (LOX) antibody (1:1000 dilution; GeneTex, Inc., Irvine, CA, USA), mouse monoclonal anti-ADAMTS1 antibody (1:500 dilution; OriGene, Rockville, MD, USA), rabbit polyclonal anti-uPAR (1:1000 dilution; GeneTex, Inc., Irvine, CA, USA), mouse monoclonal anti-α-SMA antibody (1:2000; Novus Biologicals, Littleton, CO, USA), rabbit monoclonal antibody against neuropilin-1 (NRP-1) (1:1000, clone D62C6, Cell Signaling, Danvers, MA, USA, rabbit polyclone anti-FLT1 (VEGFR1) antibody (1:1000, MyBioSource, San Diego, CA) and rabbit polyclonal anti-FAP-α antibody (1:1000, Ab207178, Abcam, Cambridge, UK).

Techniques: Western Blot

Journal: Frontiers in Molecular Biosciences

Article Title: An Extracellular Matrix-Based Signature Associated With Immune Microenvironment Predicts the Prognosis and Therapeutic Responses of Patients With Oesophageal Squamous Cell Carcinoma

doi: 10.3389/fmolb.2021.598427

Figure Lengend Snippet: The univariate Cox analysis.

Article Snippet: These microarray chips were sequentially incubated with rabbit antibodies against CST1 (Proteintech, China, 1:400), NELL2 (Proteintech, China, 1:400), ADAMTSL4 (Proteintech, China, 1:400), and ANGPTL7 (Proteintech, China, 1:400).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: An Extracellular Matrix-Based Signature Associated With Immune Microenvironment Predicts the Prognosis and Therapeutic Responses of Patients With Oesophageal Squamous Cell Carcinoma

doi: 10.3389/fmolb.2021.598427

Figure Lengend Snippet: The ECM gene model information of ESCC.

Article Snippet: These microarray chips were sequentially incubated with rabbit antibodies against CST1 (Proteintech, China, 1:400), NELL2 (Proteintech, China, 1:400), ADAMTSL4 (Proteintech, China, 1:400), and ANGPTL7 (Proteintech, China, 1:400).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: An Extracellular Matrix-Based Signature Associated With Immune Microenvironment Predicts the Prognosis and Therapeutic Responses of Patients With Oesophageal Squamous Cell Carcinoma

doi: 10.3389/fmolb.2021.598427

Figure Lengend Snippet: The differential expression of four ECM-related genes in ESCC tissues and adjacent non-tumour tissues detected by immunohistochemistry. The expression levels of CST1 (a–c) and NELL2 (d–f) were higher in ESCC tissues than in adjacent non-tumour tissues. In contrast, the expression levels of ADAMTSL4 (g–i) and ANGPTL7 (j–l) were lower in ESCC tissues (e,g) than in adjacent non-tumour tissues.

Article Snippet: These microarray chips were sequentially incubated with rabbit antibodies against CST1 (Proteintech, China, 1:400), NELL2 (Proteintech, China, 1:400), ADAMTSL4 (Proteintech, China, 1:400), and ANGPTL7 (Proteintech, China, 1:400).

Techniques: Quantitative Proteomics, Immunohistochemistry, Expressing

Journal: Frontiers in Molecular Biosciences

Article Title: An Extracellular Matrix-Based Signature Associated With Immune Microenvironment Predicts the Prognosis and Therapeutic Responses of Patients With Oesophageal Squamous Cell Carcinoma

doi: 10.3389/fmolb.2021.598427

Figure Lengend Snippet: Different expression levels of CST1, NELL2, ADAMTSL4, and ANGPTL7 proteins in the ESCC cell lines Eca109 and KYSE150 and the normal cell line Het-1A detected by western blotting.

Article Snippet: These microarray chips were sequentially incubated with rabbit antibodies against CST1 (Proteintech, China, 1:400), NELL2 (Proteintech, China, 1:400), ADAMTSL4 (Proteintech, China, 1:400), and ANGPTL7 (Proteintech, China, 1:400).

Techniques: Expressing, Western Blot