ADAMTS5 Antibody PA5-27165 from Thermo Fisher Scientific

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Yonsei medical journal
July 01 2021

Fig 1: miRNA-495 is upregulated during osteoarthritis progression in an in vitro model generated using the human normal chondrocyte cell line TC28a2. (A) TC28a2 cells were seeded at 1×105 cells per well in 24-well culture plates. The cells were treated with 10 ng/mL of IL-1ß or with no IL-1ß as a control. Safranin O staining was performed to detect glycosaminoglycans (GAGs). Stained cells were destained with 10% cetylpyridinium for quantitative analysis. Absorbance was measured at 490 nm. *p<0.05 compared to the control groups (n=3 experimental replicates). (B) The graphs represent the results of quantitative real-time polymerase chain reaction (qRT-PCR) using RNA extracted from TC28a2 cells that were treated with 10 ng/mL of IL-1ß or with no IL-1ß as a control (n=3 experimental replicates). The expression levels of all transcripts tested were normalized to those of 18S rRNA. The data are presented as means±SD. *p<0.05 compared to the IL-1ß (-) groups. (C) Protein levels of SOX9, COL2A1, COX-2, ADAMTS5, and MMP13 were analyzed in TC28a2 cells using Western blotting, and the proteins were isolated on days 0, 1, 3, and 5 after treatment with 10 ng/mL of IL-1ß. The graphs represent the average values of band intensity for each blot, and each blot was normalized to the band intensity value of ß-actin (n=3 experimental replicates). The data are presented as means±SD [COX-2 (*p<0.05 vs. Day 1), ADAMTS5 (*p<0.05 vs. Day 3), SOX9 and COL2A1 (*p<0.05 vs. Day 0)]. (D) The graphs represent the results of qRT-PCR using RNA extracted from TC28a2 cells treated with 10 ng/mL of IL-1ß or with no IL-1ß as a control (n=3 experimental replicates). The expression levels of miR-495 were normalized to those of U6. The data are presented as means±SD. *p<0.05 vs. Day 0. IL-1ß, interleukin-1ß; SFO, safranin O; CV, crystal violet; ND, not detectable.
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Oxidative medicine and cellular longevity
January 01 2022

Fig 2: BMSC-EVs enhance the survival of NPCs and inhibit ECM degradation in NPCs. (a) Characterization of BMSC-EVs by TEM observation (100 nm). (b) NanoSight analysis of the size of BMSC-EVs in diameter. (c) Western blot of EV-specific surface markers CD63, calnexin, and TSG101 in BMSC-EVs, *P < 0.05 vs. the EV group. (d) NPC uptake of PKH67-labeled BMSC-EVs by NPCs (25 µm). (e) CCK-8 assay for viability of 50 µM tBHP-exposed NPCs for 12 h following coculture with BMSC-EVs. (f) EdU staining for proliferation of 50 µM tBHP-exposed NPCs for 12 h following coculture with BMSC-EVs. (g) Flow cytometric analysis of apoptosis of 50 µM tBHP-exposed NPCs for 12 h following coculture with BMSC-EVs. (h) Immunofluorescence detection of MMP13, ADAMTS 5, COL II, and aggrecan proteins in 50 µM tBHP-exposed NPCs for 12 h following coculture with BMSC-EVs. (i) The ROS production was examined using DCF-DA kit in 50 µM tBHP-exposed NPCs for 12 h following coculture with BMSC-EVs (25 µm). (j) Lipid peroxidation determined by C11-BODIPY581/591 in 50 µM tBHP-exposed NPCs for 12 h following coculture with BMSC-EVs. (k) The total free iron in 50 µM tBHP-exposed NPCs for 12 h following coculture with BMSC-EVs. *P < 0.05 vs. the tBHP group. Cell experiments were conducted three times independently.
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Oxidative medicine and cellular longevity
January 01 2022

Fig 3: BMSC-EVs stimulated the survival of NPCs and inhibit ECM degradation in NPCs by transferring circ_0050205. (a) RT-qPCR analysis of circ_0050205 expression in the NP tissues of IDD mice (n = 8) and sham-operated mice (n = 8), *P < 0.05 vs. sham-operated mice. (b) RT-qPCR analysis of circ_0050205 expression in tBHP-exposed NPCs, *P < 0.05 vs. tBHP-0 µM. (c) Transfer of circ_0050205-Cy3 from BMSCs into NPCs via EVs (25 µm). (d) RT-qPCR analysis of circ_0050205 expression in tBHP-exposed NPCs cocultured with BMSC-EVs, *P < 0.05 vs. the tBHP group. (e) RT-qPCR analysis of circ_0050205 expression in EVs derived from circ_0050205-overexpressing BMSCs, *P < 0.05 vs. vector. (f) RT-qPCR analysis of circ_0050205 expression in tBHP-exposed NPCs cocultured with EVs derived from circ_0050205-overexpressing BMSCs, *P < 0.05 vs. tBHP + BMSC-EVs + vector. (g) CCK-8 assay for the viability of tBHP-exposed NPCs cocultured with EVs circ_0050205-overexpressing BMSCs, *P < 0.05 vs. tBHP + BMSC-EVs + vector. (h) EdU staining for proliferation of tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs, *P < 0.05 vs. tBHP + BMSC-EVs + vector. (i) Flow cytometric analysis of apoptosis of tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs, *P < 0.05 vs. tBHP + BMSC-EVs + vector. (j) Immunofluorescence detection of MMP13, ADAMTS 5, COL II, and aggrecan proteins in tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs (25 µm). (k) The ROS production was examined using the DCF-DA kit in tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs, *P < 0.05 vs. tBHP + BMSC-EVs + Vector. (l) Lipid peroxidation determined by C11-BODIPY581/591 in tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs. (m) The total free iron in tBHP-exposed NPCs cocultured with EVs from circ_0050205-overexpressing BMSCs, *P < 0.05 vs. tBHP + BMSC-EVs + vector. *P < 0.05. Cell experiments were conducted three times independently.
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Oxidative medicine and cellular longevity
January 01 2022

Fig 4: circ_0050205 in BMSC-EVs contributes to the survival of NPCs and inhibits ECM degradation in NPCs via the miR-665/GPX4 axis. (a) GPX4 expression in tBHP-exposed NPCs cocultured with BMSC-EVs-LV-circ_0050205 + sh-GPX4. (b) CCK-8 assay for viability of tBHP-exposed NPCs cocultured with BMSC-EVs-LV-circ_0050205 + sh-GPX4. (c) EdU staining for proliferation of tBHP-exposed NPCs cocultured with BMSC-EVs-LV-circ_0050205 + sh-GPX4. (d) Flow cytometric analysis of apoptosis of tBHP-exposed NPCs cocultured with BMSC-EVs-LV-circ_0050205 + sh-GPX4. (e) Immunofluorescence detection of MMP13, ADAMTS 5, COL II, and aggrecan proteins in tBHP-exposed NPCs cocultured with BMSC-EVs-LV-circ_0050205 + sh-GPX4 (25 µm). (f) The ROS production was examined using DCF-DA kit in tBHP-exposed NPCs cocultured with BMSC-EVs-LV-circ_0050205 + sh-GPX4. (g) Lipid peroxidation determined by C11-BODIPY581/591 in tBHP-exposed NPCs cocultured with BMSC-EVs-LV-circ_0050205 + sh-GPX4. (h) The total free iron in tBHP-exposed NPCs cocultured with BMSC-EVs-LV-circ_0050205 + sh-GPX4. *P < 0.05 vs. tBHP + BMSC-EVs + LV-circ_0050205 + sh-NC. Cell experiments were conducted three times independently.
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Oxidative medicine and cellular longevity
January 01 2022

Fig 5: circ_0050205 in BMSC-EVs attenuates IDD progression by regulating the miR-665/GPX4 axis. A total of 40 C57BL/6J male mice were randomly assigned into the sham group, IDD group, BMSC-EV + vector group, BMSC-EV + LV-circ_0050205 + sh-NC group, and BMSC-EV + LV-circ_0050205 + sh-GPX4 group, with 8 mice per group. (a) X-ray examination of degenerative characteristics of IDD mice and DHI (%). (b) Histological examinations of mouse NP tissues by HE staining and oil red O staining (50 µm). (c) Histological scores. (d) Immunohistochemical staining of COL II, aggrecan, and GPX4 proteins in mouse NP tissues. (e) Immunohistochemical staining of MMP13 and ADAMTS 5 proteins in mouse NP tissues. (f) The total free iron in mouse NP tissues. (g) RT-qPCR analysis of circ_0050205, miR-665, and GPX4 expression in mouse NP tissues. *P < 0.05 vs. sham, #P < 0.05 vs. IDD, &P < 0.05 vs. BMSC-EVs + vector, $P < 0.05 vs. BMSC-EVs + LV-circ_0050205 + sh-NC.

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ADAMTS5 Antibody

The ADAMTS5 Antibody from Thermo Fisher Scientific is a Rabbit Polyclonal antibody to ADAMTS5. This antibody recognizes Homo Sapiens, Human, and Mouse antigen. The ADAMTS5 Antibody has been validated for the following applications: Immunohistochemistry, Immunohistochemistry - fixed, and Western Blot.

Yonsei medical journal

Oxidative medicine and cellular longevity

Oxidative medicine and cellular longevity

Oxidative medicine and cellular longevity

Oxidative medicine and cellular longevity

ADAMTS5 Antibody from Thermo Fisher Scientific

Thermo Fisher Scientific

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ADAMTS5 Antibody from Thermo Fisher Scientific

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