Cell Culture:Article Title: SP1‑mediated ADAMTS5 transcription promotes IL‑1β‑induced chondrocyte injury via Wnt/β‑catenin pathway in osteoarthritis
Article Snippet: .. Human chondrocytes (CHON-001; American Type Culture Collection) were cultured in DMEM containing 0.1 mg/ml geneticin (G-418; Gibco; Thermo Fisher Scientific, Inc.) and 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C with 5% CO 2 . .. CHON-001 cells were stimulated with different concentrations (0, 5, 10 and 15 ng/ml) of IL-1β at 37°C for 24 h to establish the OA cell model. For transfection, cells were transfected with 50 nM ADAMTS5 siRNA [si-ADAMTS5, forward (F) 5′-AAAAUGUUUGGAUUCGUGCUC-3′; reverse (R) 5′-GCACGAAUCCAAACAUUUUCC-3′], 50 nM SP1 siRNA (si-SP1, F 5′-ACUUGAUACUGAAUAUUAGGC-3′; R 5′-CUAAUAUUCAGUAUCAAGUAA-3′), 4.0 μg pcDNA3.1 ADAMTS5 overexpression vector (F 5′-AAAGGGGAGAATCTGCCTGC-3′; R 5′-CCAAGATCCCCAGTTGCCAT-3′), 4.0 μg pcDNA3.1 SP1 overexpression vector (F 5′-GTCCGCCCTCTGACCAAG-3′; R 5′-AAGGCACCACCACCATTACC-3′) or negative controls (si-NC, F 5′-GGAGUAGGGAGCAAACCUAUAGGAA-3′, R 5′-UUCCUAUAGGUUUGCUCCCUACUCC-3′; pcDNA3.1, F 5′-CTAGAGAACCCACTGCTTAC-3′, R 5′-TAGAAGGCACAGTCGAGG-3′) using Lipofectamine ® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 24 h. All siRNAs and pcDNAs were purchased from Guangzhou RiboBio Co., Ltd. After 24 h post-transfection, cells were treated with 10 ng/ml IL-1β for 24 h at 37°C.
Article Title: SP1‑mediated ADAMTS5 transcription promotes IL‑1β‑induced chondrocyte injury via Wnt/β‑catenin pathway in osteoarthritis.
Article Snippet: .. Human chondrocytes (CHON‐001; American Type Culture Collection) were cultured in DMEM containing 0.1 mg/ml geneticin (G‐418; Gibco; Thermo Fisher Scientific, Inc.) and 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37 ̊C with 5% CO2. .. CHON‐001 cells were stimulated with different concentrations (0, 5, 10 and 15 ng/ml) of IL‐1β at 37 ̊C for 24 h to establish the OA cell model. For transfection, cells were transfected with 50 nM ADAMTS5 siRNA [si‐ADAMTS5, forward (F) 5'‐AAA AUG UUU GGA UUC GUG CUC‐3'; reverse (R) 5'‐GCA CGA AUC CAA ACA UUU UCC‐3'], 50 nM SP1 siRNA (si‐SP1, F 5'‐ACU UGA UAC UGA AUA UUA GGC‐3'; R 5'‐CUA AUA UUC AGU AUC AAG UAA‐3'), 4.0 μg pcDNA3.1 ADAMTS5 overex‐ pression vector (F 5'‐AAA GGG GAG AAT CTG CCT GC‐3'; R 5'‐CCA AGA TCC CCA GTT GCC AT‐3'), 4.0 μg pcDNA3.1 SP1 overexpression vector (F 5'‐GTC CGC CCT CTG ACC AAG‐3'; R 5'‐AAG GCA CCA CCA CCA TTA CC‐3') or negative controls (si‐NC, F 5'‐GGA GUA GGG AGC AAA CCU AUA GGA A‐3', R 5'‐UUC CUA UAG GUU UGC UCC CUA CUC C‐3'; pcDNA3.1, F 5'‐CTA GAG AAC CCA CTG CTT AC‐3', R 5'‐TAG AAG GCA CAG TCG AGG‐3') using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) at 37 ̊C for 24 h. All siRNAs and pcDNAs were purchased from Guangzhou RiboBio Co., Ltd. After 24 h post‐transfection, cells were treated with 10 ng/ml il‐1β for 24 h at 37 ̊C.
Article Title: Circular RNA circ_0020014 contributes to osteoarthritis progression via miR-613/ADAMTS5 axis
Article Snippet: .. Human chondrocytes (CHON-001; American type culture collection, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Thermo, Waltham, MA, USA) containing 10% (v/v) fetal bovine serum (GE Healthcare, Logan, UT, USA) at 37°C in a humidified incubator with 5% CO 2 . .. For IL-1β treatment, chondrocytes CHON-001 were hatched with increasing concentrations of IL-1β (Invitrogen, Carlsbad, CA, USA) for 24 hours or incubated with 10 ng/mL of IL-1β for different time.
Modification:
Article Title: Circular RNA circ_0020014 contributes to osteoarthritis progression via miR-613/ADAMTS5 axis
Article Snippet: .. Human chondrocytes (CHON-001; American type culture collection, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Thermo, Waltham, MA, USA) containing 10% (v/v) fetal bovine serum (GE Healthcare, Logan, UT, USA) at 37°C in a humidified incubator with 5% CO 2 . .. For IL-1β treatment, chondrocytes CHON-001 were hatched with increasing concentrations of IL-1β (Invitrogen, Carlsbad, CA, USA) for 24 hours or incubated with 10 ng/mL of IL-1β for different time.
Incubation:
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