Journal: Osteoarthritis and Cartilage Open

Article Title: Mild treadmill exercise inhibits cartilage degeneration via macrophages in an osteoarthritis mouse model

doi: 10.1016/j.ocarto.2023.100359

Figure Lengend Snippet: Treadmill exercise affects cartilage degeneration. (A) Histological images of articular cartilage stained with Safranin-O Fast Green and immunohistochemical staining for Gremlin-1, MMP-13, and ADAMTs4 (Scale bars: 50 ​μm). (B) Scoring results by OARSI score to assess cartilage degeneration and positive cell rates for Gremlin-1, MMP-13, and ADAMTs4 (n ​= ​8, in each group. One-Way ANOVA, post-hoc Tukey). Data are presented as means and 95% confidence interval. In mild-ex group, OARSI score, MMP-13, and ADAMTS4 positive cell rate were suppressed. Positive cell rate for gremlin-1 increased in an exercise intensity-dependent manner.

Article Snippet: Anti-gremlin-1 (ab231065; Abcam plc．MA，USA，1:200), anti-MMP-13 (ab39012; Abcam plc．MA，USA，1:250), anti-ADAMTS4 (bs-4191R; Bioss Antibodies Co., Ltd. MA, USA, 1:100), anti-CD68 (ab125212; Abcam plc.

Techniques: Staining, Immunohistochemical staining

Journal: Osteoarthritis and Cartilage Open

Article Title: Mild treadmill exercise inhibits cartilage degeneration via macrophages in an osteoarthritis mouse model

doi: 10.1016/j.ocarto.2023.100359

Figure Lengend Snippet: Treadmill exercise and clodronate liposomes affect cartilage degeneration. (A) Histological images of articular cartilage stained with Safranin-O Fast Green and immunohistochemical staining for Gremlin-1, MMP-13, and ADAMTs4 (Scale bars: 50 ​μm). (B) Scoring results by OARSI score to assess cartilage degeneration and positive cell rates for Gremlin-1, MMP-13, and ADAMTs4 (n ​= ​8, in each group. One-Way ANOVA, post-hoc Tukey). Data are presented as means and 95% confidence interval. Ex and MΦ-de group showed lower OARSI scores and lower MMP-13 positive cell rates. Gremlin-1 positive cell rate increased in the exercise group.

Article Snippet: Anti-gremlin-1 (ab231065; Abcam plc．MA，USA，1:200), anti-MMP-13 (ab39012; Abcam plc．MA，USA，1:250), anti-ADAMTS4 (bs-4191R; Bioss Antibodies Co., Ltd. MA, USA, 1:100), anti-CD68 (ab125212; Abcam plc.

Techniques: Liposomes, Staining, Immunohistochemical 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. 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: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Partially redundant functions of Adamts1 and Adamts4 in the perinatal development of the renal medulla.

doi: 10.1002/dvdy.22662

Figure Lengend Snippet: Fig. 1. Targeting of Adamts4 in mice. A: Illustration of the targeting strategy used to generate the Adamts4flox and Adamts4 alleles. The target- ing strategy introduced a unique AflII restriction site, resulting in a novel 9.7-kb restriction fragment that was detectable by Southern blotting using the indicated probe. Neo, neomycin resistance cassette; DTa, diphtheria toxin a-chain. Exons are illustrated as boxes and numbered with roman numerals. B: Generation of embryonic stem cell lines heterozygous for the Adamts4flox allele. The presence of the targeted allele was detected as a novel AflII restriction fragment by Southern blotting (clone BG3). C: Representative PCR genotyping analyses showing the relative migration of PCR products derived from the Adamts4 wild-type, floxed, and null alleles. The genotypes of the animals are indicated above each lane. D: RT- PCR analysis of Adamts4 expression in kidneys from mice of the indicated genotypes. Results confirm the loss of Adamts4 mRNA in Adamts4/

Article Snippet: The primary antibody against ADAMTS4 (Aviva Systems Biology, San Diego, CA, catalog no. ARP41556) was used at a dilution of 1:200 for 2 hr at room temperature.

Techniques: Southern Blot, Migration, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Partially redundant functions of Adamts1 and Adamts4 in the perinatal development of the renal medulla.

doi: 10.1002/dvdy.22662

Figure Lengend Snippet: Fig. 3. Perinatal onset of the renal pheno- type in Adamts1/;Adamts4/ mice. Photo- micrographs show kidneys from 15.5dpc (A, B) and 17.5dpc (C, D) wild-type and Adamts1/;Adamts4/ mice, all of which are morphologically normal. For both genotypes and embryonic stages, n ¼ 3 animals (6 kid- neys) were examined. Adult kidneys from wild-type (E) and Adamts1/;Adamts4/ (F) mice showed a markedly thinned medulla and pelvis dilation in the Adamts1/;Adamts4/

Article Snippet: The primary antibody against ADAMTS4 (Aviva Systems Biology, San Diego, CA, catalog no. ARP41556) was used at a dilution of 1:200 for 2 hr at room temperature.

Techniques:

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Partially redundant functions of Adamts1 and Adamts4 in the perinatal development of the renal medulla.

doi: 10.1002/dvdy.22662

Figure Lengend Snippet: Fig. 4. Adamts1, Adamts4, Vcan, and Acan expression in perinatal kidneys. Real-time RTPCR timecourse analysis of Adamts1 (A), Adamts4 (B), and Vcan (C) expression in 15.5-dpc, 17.5- dpc, 1-d, and 7-d wild-type mouse kidneys. Vcan was more highly expressed at 15.5 dpc com- pared with 17.5 dpc, 1 d, and 7 d in the perinatal kidney (P < 0.05). D: RT-PCR analysis of Acan expression demonstrates the presence of Acan mRNA in the newborn mouse kidney, albeit at low levels compared to brain and cartilage.

Article Snippet: The primary antibody against ADAMTS4 (Aviva Systems Biology, San Diego, CA, catalog no. ARP41556) was used at a dilution of 1:200 for 2 hr at room temperature.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Partially redundant functions of Adamts1 and Adamts4 in the perinatal development of the renal medulla.

doi: 10.1002/dvdy.22662

Figure Lengend Snippet: Fig. 5. Immunohistochemical localization of ADAMTS4 expression in newborn and 3-month-old adult wild-type mouse kidneys. A: Negative control for newborn kidney. B: ADAMTS4 was local- ized mainly to the developing distal and proximal tubules in newborn mice. C,E: Negative con- trol for adult kidney. D,F: ADAMTS4 localized predominantly to the distal and proximal tubules (arrows), with minimal staining in the glomeruli and medulla. Staining in the area of the renal capsule was frequently observed in both control (IgG) and experimental samples, and is believed to be an edge artefact. M, medulla; CT, convoluted tubule; G, glomerulus.

Article Snippet: The primary antibody against ADAMTS4 (Aviva Systems Biology, San Diego, CA, catalog no. ARP41556) was used at a dilution of 1:200 for 2 hr at room temperature.

Techniques: Immunohistochemical staining, Expressing, Negative Control, Staining, Control

Journal: Arthritis and Rheumatism

Article Title: ADAMTS-4_v1 Is a Splice Variant of ADAMTS-4 That Is Expressed as a Protein in Human Synovium and Cleaves Aggrecan at the Interglobular Domain

doi: 10.1002/art.38102

Figure Lengend Snippet: Aggrecanase substrate activity of ADAMTS-4_v1, as assessed by SensoLyte 520 aggrecanase 1 assay. A, Aggrecanase 1 activity of an anti–FLAG M2 immunoprecipitate from ADAMTS-4_v1 transfected with HEK 293 cells that had been predigested for 1 hour at 37°C with furin (▪) or an equal amount of undigested ADAMTS-4_v1 immunoprecipitate (□), as compared to that of the recombinant truncated ADAMTS-4 (•) (supplied in the assay kit). Enzyme activity (expressed as relative fluorescence units [RFU]) was measured every 5 minutes for 1 hour. B, Western blot probed with anti–FLAG M2 monoclonal antibody from anti–FLAG M2 immunoprecipitates from ADAMTS-4_v1–transfected HEK 293 cells used in A. Shown are undigested and furin-digested ADAMTS-4_v1 prior to their use in the assay (preassay) as well as undigested and furin-digested ADAMTS-4_v1 plus recombinant truncated ADAMTS-4 (rADAMTS-4) that was harvested from the assay plate after the 1-hour course of the aggrecanase 1 assay (postassay). C, Duplicate Western blot of B probed with affinity-purified goat anti-human ADAMTS-4 antibodies (BAF4307).

Article Snippet: Primary antibodies ab28285 and ab39201 were from Abcam and BAF4307 from R&D Systems.

Techniques: Activity Assay, Transfection, Recombinant, Fluorescence, Western Blot, Affinity Purification